Cyclic motilin receptor antagonists

ABSTRACT

The present invention aims to provide cyclic peptide derivatives having motilin receptor antagonist activity and are useful as pharmaceuticals. The present invention provides compounds of general formula (1):                  
 
wherein R 1  represents an optionally substituted phenyl group or the like; R 2  represents an amino group or the like; R 3  to R 6  represent a hydrogen atom, a methyl group or the like; R 7  represents a hydrogen atom or the like; V to Z represent a carbonyl group or a methylene group; m represents an integer of 0–2; and n represents an integer of 0–3; or a hydrate or a pharmaceutically acceptable salt thereof.

TECHNICAL FIELD

The present invention relates to cyclic peptide derivatives havingmotilin receptor antagonist activity or the like and useful aspharmaceuticals.

BACKGROUND ART

Among gastrointestinal hormones, motilin is a straight peptideconsisting of 22 amino acids that is known to control gastrointestinalmotility of mammals including human. Motilin receptors have been knownto be predominantly localized in the upper gastrointestinal tract suchas stomach and duodenum, and recently found to be also localized in thelower gastrointestinal tract such as large intestine (William et al.,Am. J. Physiol., 262, G50–G55 (1992)), showing that motilin may beinvolved in the motility of not only the upper but also the lowergastrointestinal tract.

It was reported that patients of irritable bowel syndrome showingdiarrhea conditions or patients of irritable bowel syndrome under stressshow hypermotilinemia (Preston et al., Gut, 26, 1059–1064 (1985); Fukodoet al., Tohoku J. Exp. Med., 151, 373–385 (1987)), suggesting thatincreased blood motilin may be involved in this pathology. Otherpathologies reported to be associated with hypermotilinemia includeCrohn's disease, ulcerative colitis, pancreatitis, diabetes, obesity,malabsorption syndrome, bacterial diarrhea, atrophic gastritis,postgastrectomy/enterectomy, etc. Therefore, motilin receptorantagonists may potentially improve pathologies with increased bloodmotilin such as irritable bowel syndrome.

Recently, efforts have been made to develop and research motilinreceptor antagonists, and various compounds have been reported(JP-A-7-138284, JP-A-2000-44595, etc.).

Especially, JP-A-7-138284 discloses cyclic peptide derivatives, whichare used as pharmacological tools in studies of the effect of motilin ongastrointestinal motility or development and research of pharmaceuticalsin this field of the art. However, their motilin antagonist activity isnot sufficient and it would be desirable to develop cyclic peptidederivatives having higher activity.

DISCLOSURE OF THE INVENTION

An object of the present invention is to provide cyclic peptidederivatives having motilin receptor antagonist activity and useful aspharmaceuticals.

As a result of careful studies to develop novel cyclic peptidederivatives having higher motilin receptor antagonist activity, weaccomplished the present invention on the basis of the finding thatcyclic peptide derivatives of general formula (1) have excellent motilinreceptor antagonist activity.

Accordingly, the present invention provides a compound of generalformula (1):

wherein

-   R₁ represents an optionally substituted phenyl group or an    optionally substituted heterocycle;

-   R₂ represents a hydrogen atom or an optionally substituted amino    group;

-   R₃ represents a hydrogen atom, a methyl group or an ethyl group;

-   R₄ represents a hydrogen atom, a methyl group or an ethyl group;

-   R₅ represents a hydrogen atom, a methyl group or an ethyl group;

-   R₆ represents a hydrogen atom, a methyl group, an ethyl group or    —CO—R₇;

-   R₇ represents a hydrogen atom or a straight or branched alkyl group    having 1–3 carbon atoms;

-   V represents a carbonyl group or a methylene group;

-   W represents a carbonyl group or a methylene group;

-   X represents a carbonyl group or a methylene group;

-   Y represents a carbonyl group or a methylene group;

-   Z represents a carbonyl group or a methylene group;

-   m represents a number of 0–2; and

-   n represents a number of 0–3;

-   except for the case where R₁ represents a phenyl group, and R₂    represents an amino group, and all of R₃, R₄, R₅ and R₆ represent a    hydrogen atom, and V represents a methylene group, and all of W, X,    Y and Z represent a carbonyl group, and both m and n represent 1; or

-   a hydrate or a pharmaceutically acceptable salt thereof.

-   

The present invention also provides a pharmaceutical comprising thecompound of general formula (1) or a hydrate or a pharmaceuticallyacceptable salt thereof as an active ingredient. The present inventionalso provides a motilin receptor antagonist comprising the abovecompound or a hydrate or a pharmaceutically acceptable salt thereof. Thepresent invention also provides a gastrointestinal motility inhibitorcomprising the above compound or a hydrate or a pharmaceuticallyacceptable salt thereof as an active ingredient. The present inventionalso provides a pharmaceutical for treating hypermotilinemia comprisingthe above compound or a hydrate or a pharmaceutically acceptable saltthereof as an active ingredient. The above compound or a hydrate or apharmaceutically acceptable salt thereof may also be used for preparinga pharmaceutical or a pharmaceutical composition such as motilinreceptor antagonists, gastrointestinal motility depressants andpharmaceuticals for treating hypermotilinemia.

According to another aspect of the present invention, a method fortreating hypermotilinemia with the above compound or a hydrate or apharmaceutically acceptable salt thereof is provided. Preferably, amethod for treating hypermotilinemia associated with irritable bowelsyndrome, Crohn's disease, ulcerative colitis, pancreatitis, diabetes,obesity, malabsorption syndrome, bacterial diarrhea, atrophic gastritis,postgastrectomy/enterectomy and the like is provided. For example, amethod comprising the step of administering a therapeutically effectiveamount of the above compound or a hydrate or a pharmaceuticallyacceptable salt thereof to a patient in need of such treatment isprovided.

The present invention also provides a compound of general formula (2):

wherein R₃, R₄, R₅, R₆, m and n have the same meanings as defined forgeneral formula (1) above;

-   P₂ represents a hydrogen atom or a protective group for a phenolic    hydroxyl group;-   P₃ represents a hydrogen atom or a protective group for a carboxyl    group;-   P₄ represents a hydrogen atom or a protective group for an amino    group; and-   P₅ represents a hydrogen atom or a protective group for an amino    group; or-   a hydrate or a pharmaceutically acceptable salt thereof.

The present invention also provides a compound of general formula (3):

wherein R₃, R₄, R₅, m, n, P₂, P₃ and P₅ have the same meanings asdefined for general formula (2) above;

-   R₁ has the same meaning as defined in general formula (1) above; and-   R₂′ represents a hydrogen atom or an optionally substituted    protected amino group; or-   a hydrate or a pharmaceutically acceptable salt thereof.

The present invention also provides a compound of general formula (4):

wherein R₁, R₃, R₄, R₅, R₂′, m and n have the same meanings as definedfor general formula (3) above;

-   P₆ represents a hydrogen atom or a protective group for a carboxyl    group; and-   P₇ represents a hydrogen atom or a protective group for an amino    group; or-   a hydrate or a pharmaceutically acceptable salt thereof.

In the definition of the compound of general formula (1), substituentsin the optionally substituted phenyl group for R₁ preferably includehalogen atoms and trifluoromethyl, hydroxyl, amino and nitrile groups,more preferably halogen atoms, especially fluorine atom. The phenylgroup may have one or more of these substituents which may be the sameor different. The number of substituents is preferably 1–3, morepreferably 1.

The optionally substituted phenyl group for R₁ may be phenyl,4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl, 3,4-difluorophenyl,2,3,4-trifluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 2-chlorophenyl,3,4-dichlorophenyl, 2,3,4-trichlorophenyl, 4-trifluoromethylphenyl,3-trifluoromethylphenyl, 4-hydroxyphenyl, 3-hydroxyphenyl,4-aminophenyl, 4-cyanophenyl, 3-cyanophenyl, 3-fluoro-4-hydroxyphenyl,2-fluoro-4-hydroxyphenyl or the like, preferably phenyl, 4-fluorophenyl,3-fluorophenyl, 3,4-difluorophenyl, 4-chlorophenyl, 3-chlorophenyl or3,4-dichlorophenyl, more preferably 4-fluorophenyl.

The heterocycle in the optionally substituted heterocycle for R₁ may bean aliphatic or aromatic 5 to 7-membered monocycle or fused cyclecontaining at least one hetero atoms selected from nitrogen, sulfur andoxygen atoms, specifically pyridyl, furyl, thienyl, indolyl, quinolinyl,benzofuryl, tetrahydroisoquinolyl, preferably indolyl.

Substituents in the optionally substituted heterocycle for R₁ includehydroxyl, amino, carboxyl, methoxyl, methyl, ethyl, trifluoromethyl,oxo, etc., and the heterocycle may have one or more of thesesubstituents which may be the same or different.

The optionally substituted heterocycle for R₁ is preferably 3-indolyl.

R₁ as defined above is preferably phenyl, 4-fluorophenyl,3-fluorophenyl, 3,4-difluorophenyl, 4-chlorophenyl, 3-chlorophenyl,3,4-dichlorophenyl or 3-indolyl, more preferably 4-fluorophenyl.

Substituents in the optionally substituted amino group for R₂ may be,for example, a straight or branched alkyl group having 1–3 carbon atoms,preferably methyl or ethyl. The amino group may have one or more ofthese substituents which may be the same or different.

The optionally substituted amino group for R₂ may be an amino groupwhich may be substituted by one or more the same or different straightor branched alkyl groups having 1–3 carbon atoms, such as amino,methylamino, ethylamino, dimethylamino, among which amino is especiallypreferred.

R₂ as defined above is preferably an amino group.

R₃ is preferably a hydrogen atom or a methyl group.

R₄ is preferably a hydrogen atom or a methyl group.

R₅ is preferably a hydrogen atom or a methyl group.

R₆ is preferably a hydrogen atom, a methyl group or an acetyl group.

V is preferably a methylene group.

W is preferably a carbonyl group or a methylene group.

X is preferably a carbonyl group or a methylene group.

Y is preferably a carbonyl group.

Z is preferably a carbonyl group.

m is preferably 0 or 1.

n is preferably 0, 1 or 2.

The sum of m and n is preferably 1, 2 or 3.

In a preferred compound of general formula (1), X is a methylene groupwhen R₁ a phenyl group and R₃ is a hydrogen atom.

In another preferred compound of general formula (1), R₁ is a4-fluorophenyl group, R₂ is an amino group, R₃ is a hydrogen atom or amethyl group, R₄ is a hydrogen atom or a methyl group, R₅ is a hydrogenatom or a methyl group, R₆ is a hydrogen atom, a methyl group or anacetyl group, V is a methylene group, W is a carbonyl group or amethylene group, X is a carbonyl group or a methylene group, Y is acarbonyl group, Z is a carbonyl group, m is 0 or 1, n is 0, 1 or 2 andthe sum of m and n is 1, 2 or 3. More preferably, the compound is anyone of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S,14S)-13-(2S-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane,(2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-N-methyl-1,4,8-triaza-3,14-dioxocyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)propionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9,N-dimethyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)propionamide,and(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide.

The compounds of general formulae (2)-(4) are useful as intermediatesfor preparing the compound of general formula (1). Protective groups foramino, carboxyl and phenolic hydroxyl groups as defined for thesegeneral formulae (2)–(4) include the following groups.

Protective groups for amino groups (i.e. P₄, P₅, P₇ and the protectivegroup in R₂′ which is an optionally substituted protected amino group)include functional groups known to be useful as protective groups foramino groups such as benzyloxycarbonyl, t-butoxycarbonyl,9-fluorenylmethyloxycarbonyl, allyloxycarbonyl,2,2,2-trichloroethoxycarbonyl, benzoyl, acetyl, trifluoroacetyl,benzenesulfonyl, p-toluenesulfonyl, trimethylsilyl,t-butyldimethylsilyl, benzyl and benzyloxymethyl, preferablybenzyloxycarbonyl and t-butoxycarbonyl.

Protective groups for carboxyl groups (P₃ and P₆) include functionalgroups known to be useful as protective groups for carboxyl groups suchas methyl, ethyl, t-butyl, allyl, benzyl, 2,2,2-trichloroethyl,trimethylsilyl and t-butyldimethylsilyl, preferably methyl, benzyl andt-butyl.

Protective groups for phenolic hydroxyl groups (P₂) include functionalgroups known to be useful as protective groups for phenolic hydroxylgroups such as methyl, methoxymethyl, benzyloxymethyl,2-methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl,tetrahydropyranyl, allyl, t-butyl, benzyl, 4-methoxybenzyl,2-nitrobenzyl, trimethylsilyl, t-butyldimethylsilyl, acetyl,trifluoroacetyl, benzoyl, methoxycarbonyl, 2,2,2-trichloroethoxycarbonyland t-butoxycarbonyl, preferably benzyl.

Acids forming a salt of the compound of formula (1), (2), (3) or (4)include inorganic acids such as hydrochloric acid, hydrobromic acid,hydroiodic acid, sulfuric acid and phosphoric acid, and organic acidssuch as acetic acid, oxalic acid, maleic acid, fumaric acid, citricacid, succinic acid, tartaric acid, methanesulfonic acid andtrifluoroacetic acid.

Compounds of the present invention may exist as optical isomers, andsuch individual optical isomers and mixtures thereof are all included inthe present invention.

Compounds of the present invention may also be obtained as hydrates ofthe compounds of formula (1), (2), (3) or (4).

The present invention is specifically explained below, in which aminoacids forming peptides, protective groups and reagents are sometimesabbreviated as follows. Tyr: tyrosine; Z: benzyloxycarbonyl; Boc:tert-(or t-)butoxycarbonyl; CMPI: 2-chloro-1-methylpyridinium iodide;PyCIU: chloro-N,N,N′,N′-bis(tetramethylene)formamidiniumhexafluorophosphate; PyBrop: bromotripyrrolidinophosphoniumhexafluorophosphate; BOP:benzotriazole-1-yl-oxy-tris(dimethylamino)phosphoniumhexafluorophosphate; HATU:O-(7-azabenzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate; WSCI: N-ethyl-N′-3-dimethylaminopropylcarbodiimide; DIC: N,N′-diisopropyl carbodiimide; DCC: N,N′-dicyclohexylcarbodiimide; DPPA: diphenylphosphoryl azide; CDI:1,1′-carbonyldiimidazole; HOBT: 1-hydroxybenzotriazole monohydrate; NMM:N-methylmorpholine; TEA: triethylamine; DIEA: diisopropylethylamine;DMAP: 4-dimethylaminopyridine; TFA: trifluoroacetic acid; THF:tetrahydrofuran; DMF: N,N-dimethylformamide; CH: chloroform; MC:methylene chloride; M: methanol; N: concentrated aqueous ammonia; EA:ethyl acetate; H: n-hexane.

PREFERRED EMBODIMENTS OF THE INVENTION

The present application claims priority based on Japanese PatentApplication No. 2000-253950, the disclosure of which is incorporatedherein as reference in its entirety.

Compounds of the present invention can be prepared by solid phase orliquid phase processes. In the case of solid phase processes, they canbe prepared by using an automatic organic synthesizer or manually.

Compounds of the present invention can be prepared by the process ofscheme 1, 2 or 3 described below, which may be modified in part asappropriate to suit the desired compound, starting from a knowncompound. Compounds of the present invention can also be obtained byappropriately adapting the specific processes described in the examplesbelow.

Scheme 1

Compounds of the present invention wherein V represents a methylenegroup and W, X, Y and Z represent a carbonyl group can be preparedaccording to scheme 1 shown below using reagents having desired groups.

wherein P₁, P₄ and P₅ represent a protective group for an amino group,P₂ represents a hydrogen atom or a protective group for a phenolichydroxyl group, P₃ represents a protective group for a carboxyl group,R₂′ represents a hydrogen atom or an optionally substituted protectedamino group and the other symbols have the same meanings as defined inclaim 1.

In scheme 1, conversion from compound 1 (described in Example 1(1) ofJPA 2000-44595) into compound 2 can be performed by protecting the aminogroup of compound 1 and then, if desired, protecting the phenol group.Specifically, compound 2 wherein P₂ is a hydrogen atom can be obtainedby reacting compound 1 with a protective group-introducing reagent suchas benzyloxycarbonyl chloride or di-t-butyl dicarbonate in a mixedsolvent of dioxane and water in the presence of a base such as sodiumcarbonate with ice cooling or at room temperature. Compound 2 wherein P₂is a protective group (especially, benzyl) can be obtained by furtherreacting the compound with benzyl chloride or the like in DMF or THF inthe presence of a base such as sodium hydride with ice cooling.

Conversion from compound 2 into compound 3 can be performed by treatingthe methyl ester group of compound 2 with a base to hydrolyze it.Specifically, compound 3 can be obtained by stirring compound 2 with abase such as sodium hydroxide or lithium hydroxide in a mixed solvent ofmethanol and water or dioxane and water with ice cooling or at roomtemperature.

Conversion from compound 3 into compound 4 can be performed by reactingcompound 3 with a methylating agent or an ethylating agent in thepresence of a base. Specifically, compound 4 can be obtained by reactingcompound 3 with methyl iodide, dimethyl sulfate or ethyl iodide or thelike in DMF or THF or a mixed solvent of DMF and THF in the presence ofa base such as sodium hydride with ice cooling. The phenolic hydroxylgroup here is preferably protected by P₂. This methylation or ethylationreaction is unnecessary for compounds wherein R₄ is a hydrogen atom.

Conversion from compound 4 into compound 6 is performed by reactingcompound 4 with compound 5. This reaction can be performed by condensingthe carboxyl group of compound 4 with the amino group of compound 5using a known conventional condensing reagent. Specifically, compound 6can be obtained by stirring a mixture of compound 4, compound 5 and oneof condensing agents such as CMPI, BOP, PyCIU, PyBrop, HATU, DCC, DIC,WSCI, DPPA and CDI in a reaction solvent such as THF or DMF or methylenechloride. The reaction here is often preferably promoted by adding abase such as TEA, DIEA, NMM, pyridine and DMAP or adding an additivesuch as HOBT as appropriate. Compound 5 may be commercially available.If not commercially available, compound 5 wherein R₅ is a hydrogen atomcan be prepared by esterifying a suitable α-, β-, γ- or δ-amino acid.Compound 5 wherein R₅ is a methyl or ethyl group can be obtained byprotecting the amino group of a suitable α-, β-, γ- or δ-amino acidester with a benzyloxycarbonyl group or a t-butoxycarbonyl group in thesame manner as described for the conversion from compound 1 intocompound 2, methylating or ethylating the amino acid ester in the samemanner as described for the conversion from compound 3 into compound 4,and then eliminating the amino-protecting group by a conventionalmethod.

Conversion from compound 6 into compound 7 can be performed byeliminating the amino-protecting group P₁ of compound 6. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₁ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

Conversion from compound 7 into compound 9 is performed by reactingcompound 7 with compound 8. This reaction can be performed by condensingthe amino group of compound 7 with the carboxyl group of compound 8using a known conventional condensing reagent. Specifically, compound 9can be obtained by the same procedure as described for the conversionfrom compound 4 into compound 6. Compound 8 may be commerciallyavailable. If not commercially available, it can be obtained bysimultaneously methylating both protected amino groups of a precursor ofcompound 8 wherein the α-amino group and the pendant amino groupcorresponding to R₃ and R₆ are protected by P₄ and P₅ respectively (P₄and P₅ are preferably different protective groups) in the same manner asdescribed for the conversion from compound 3 into compound 4. Compounds8 other than those wherein both R₃ and R₆ represent a methyl group canalso be prepared by appropriately adapting this procedure.

Conversion from compound 9 into compound 10 can be performed byeliminating the amino-protecting group P₄ of compound 9. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₄ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

Conversion from compound 10 into compound 12 is performed by reactingcompound 10 with compound 11. This reaction can be performed bycondensing the amino group of compound 10 with the carboxyl group ofcompound 11 using a known conventional condensing reagent. Specifically,compound 12 can be obtained by the same procedure as described for theconversion from compound 4 into compound 6. Compound 11 may becommercially available. If not commercially available, compound 11wherein R₂′ is a protected methylamino group can be obtained bymethylating a precursor of compound 11 wherein the group correspondingto R₂′ is a protected amino group in the same manner as described forthe conversion from compound 3 into compound 4. Compounds 11 other thanthose wherein the group corresponding to R₂′ is a protected amino groupcan also be prepared by appropriately adapting this procedure.

Conversion from compound 12 into compound 13 can be performed bysimultaneously or stepwise eliminating the amino-protecting group P₅,carboxyl-protecting group P₃ and phenol-protecting group P₂ of compound12. A specific deprotecting method comprises hydrogenolysis using apalladium catalyst when P₅ is a benzyloxycarbonyl group, P₃ is a benzylgroup and P₂ is a benzyl group or treatment with an acid such as TFAwhen P₅ is a t-butoxycarbonyl group and P₃ is t-butyl or alkalihydrolysis when P₃ is a methyl group, for example.

Conversion from compound 13 into compound 14 can be performed byintramolecular cyclization of the amino group and the carboxyl group ofcompound 13 using a condensing reagent known to be useful for macrocycleformation reaction. Specifically, compound 14 can be obtained bystirring compound 13 at room temperature under dilute conditions (at aconcentration of compound 13 of about 0.005–0.02 M, preferably 0.01 M)using DMF, BOP and pyridine as reaction solvent, condensing agent andbase, respectively.

When R₂′ of compound 14 is an optionally substituted protected aminogroup, conversion from compound 14 into compound 15 can be performed bydeprotecting the amino group. A specific method comprises hydrogenolysisusing a palladium catalyst when the amino-protecting group of R₂′ is abenzyloxycarbonyl group or treatment with an acid such as TFA when it isa t-butoxycarbonyl group. When R₂′ is a hydrogen atom, compound 14 is adesired compound.

Scheme 2

Compounds of the present invention wherein V and X represent a methylenegroup and W, Y and Z represent a carbonyl group can be preparedaccording to scheme 2 shown below using reagents having desired groups.

wherein P₄ and P₅ represent a protective group for an amino group, P₂represents a hydrogen atom or a protective group for a phenolic hydroxylgroup, P₃ represents a protective group for a carboxyl group, R₂′represents a hydrogen atom or an optionally substituted protected aminogroup when R₂ of the desired compound represents an optionallysubstituted amino group, R₇ represents a functional group capable ofreacting with an amino group to form a bond such as formyl, —CH₂Halwhere Hal represents a halogen atom such as a chlorine, bromine oriodine atom, —CH₂OSO₂R where R represents a methyl, trifluoromethyl,tosyl or the like group, and the other symbols have the same meanings asdefined in claim 1.

Conversion from compound 9 into compound 16 can be performed bysimultaneously or stepwise eliminating the amino-protecting group P₅,carboxyl-protecting group P₃ and phenol-protecting group P₂ of compound9. A specific deprotecting method comprises hydrogenolysis using apalladium catalyst when P₅ is a benzyloxycarbonyl group, P₃ is a benzylgroup and P₂ is a benzyl group or treatment with an acid such as TFAwhen P₅ is a t-butoxycarbonyl group and P₃ is t-butyl or alkalihydrolysis when P₃ is a methyl group, for example.

Conversion from compound 16 into compound 17 can be performed byintramolecular cyclization of the amino group and the carboxyl group ofcompound 16 using a condensing reagent known to be useful for macrocycleformation reaction. Specifically, compound 17 can be obtained bystirring compound 16 at room temperature under dilute conditions (at aconcentration of compound 16 of about 0.005–0.02 M, preferably 0.01 M)using DMF, BOP and pyridine as reaction solvent, condensing agent andbase, respectively.

Conversion from compound 17 into compound 18 can be performed byeliminating the amino-protecting group P₄ of compound 17. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₄ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

Conversion from compound 18 into compound 20 is performed by reactingcompound 18 with compound 19. When R₇ of compound 19 is a formyl group,this reaction can be performed by reductively forming a bond betweenthis formyl group and the amino group of compound 18, specificallystirring a mixture of compound 18 and compound 19 with sodiumcyanoborohydride and acetic acid in a reaction solvent such as methanolor acetonitrile, for example. When R₇ of compound 19 is —CH₂Hal or—CH₂OSO₂R or the like, the reaction can be performed by alkylating theamino group of compound 18. Compound 19 here wherein R₇ is a formylgroup and R₂′ is an optionally substituted protected amino group, forexample, can be obtained by condensing the carboxyl group of compound 11used in scheme 1 wherein R₂′ represents an optionally substitutedprotected amino group with N,O-dimethylhydroxylamine to convert it intoan N-methoxy-N-methylcarbamoyl group and then reducing theN-methoxy-N-methylcarbamoyl group with lithium aluminium hydride, forexample.

When R₂′ of compound 20 is an optionally substituted protected aminogroup, conversion from compound 20 into compound 21 can be performed bydeprotecting the amino group, specifically catalytic hydrogenation whenthe amino-protecting group of R₂′ is a benzyloxycarbonyl group ortreatment with an acid such as TFA when it is a t-butoxycarbonyl group.When R₂′ is a hydrogen atom, compound 21 is a desired compound.

Scheme 3

Compounds of the present invention wherein R₆ represents an acetylgroup, V and W represent a methylene group and X, Y and Z represent acarbonyl group can be prepared according to scheme 3 shown below usingreagents having desired groups.

wherein P₁, P₄, P₅ and P₇ represent a protective group for an aminogroup, P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group, P₆ represents a protective group for a carboxylgroup, R₂′ represents a hydrogen atom or an optionally substitutedprotected amino group when R₂ of the desired compound represents anoptionally substituted amino group, R₈ represents a functional groupcapable of reacting with an amino group to form a bond such as formyl,—CH₂Hal where Hal represents a halogen atom such as a chlorine, bromineor iodine atom, —CH₂OSO₂R where R represents a methyl, trifluoromethyl,tosyl or the like group, and the other symbols have the same meanings asdefined in claim 1.

Conversion from compound 4 into compound 22 can be performed bysuccessively esterifying the carboxyl group of compound 4 with P₆ andeliminating the amino-protecting group P₁ and phenol-protecting groupP₂. Specifically, the carboxyl group can be esterified with P₆ into abenzyl ester by condensation reaction with benzyl alcohol or into at-butyl ester by a reaction with isobutene and an acid or into a methylester by a reaction with methanol or methylation reaction withdiazomethane. The amino-protecting group can be eliminated by treatmentwith an acid such as TFA when it is a t-butoxycarboxy group orhydrogenolysis using a palladium catalyst when it is a benzyloxycarboxygroup. When the phenol-protecting group P₂ is a t-butyl or benzyl group,P₁ and P₂ can be simultaneously eliminated. Compound 22 wherein R₄ is ahydrogen atom and P₆ is a methyl group is compound 1.

Conversion from compound 22 (or compound 1) into compound 24 isperformed by reacting compound 22 (or compound 1) with compound 23. Thisreaction can be performed by condensing the amino group of compound 22(or compound 1) with the carboxyl group of compound 23 using a knownconventional condensing reagent. Specifically, compound 24 can beobtained by the same procedure as described for the conversion fromcompound 4 into compound 6 in scheme 1. Compound 23 may be commerciallyavailable. If not commercially available, it can be obtained by adaptingthe process for compound 8 described in scheme 1. Compound 23 wherein mis 1, R₃ is a hydrogen atom or a methyl group, any one of P₄ and P₅ is at-butoxycarbonyl group and the other is a benzyloxycarbonyl group iscommercially available.

Conversion from compound 24 into compound 25 can be performed byeliminating the amino-protecting group P₄ of compound 24. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₄ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

Conversion from compound 25 into compound 26 is performed by reactingcompound 25 with compound 11 used in scheme 1. This reaction can beperformed by condensing the amino group of compound 25 with the carboxylgroup of compound 11 used in scheme 1 using a known conventionalcondensing reagent. Specifically, compound 26 can be obtained by thesame procedure as described for the conversion from compound 4 intocompound 6 in scheme 1.

Conversion from compound 26 into compound 27 can be performed byeliminating the amino-protecting group P₅ of compound 26. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₅ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

Conversion from compound 27 into compound 29 is performed by reactingcompound 27 with compound 28. When R₈ of compound 28 is a formyl group,this reaction can be performed by reductively forming a bond with theamino group of compound 27, specifically stirring a mixture of compound27 and compound 28 with sodium cyanoborohydride and acetic acid in areaction solvent such as methanol or acetonitrile, for example. When R₇of compound 28 is —CH₂Hal or —CH₂OSO₂R or the like, the reaction can beperformed by alkylating the amino group of compound 27. Compound 28 herewherein R₈ is a formyl group, for example, can be obtained by reducingthe carboxyl group of a precursor of compound 28 wherein the groupcorresponding to R₈ is a carboxyl group with borane or the like into ahydroxymethyl group and then oxidizing the hydroxymethyl group into aformyl group by the Swern method or the like, for example.Alternatively, it can also be obtained by condensing the carboxyl groupof a precursor of compound 28 wherein the group corresponding to R₈ is acarboxyl group with N,O-dimethylhydroxylamine to convert it into anN-methoxy-N-methylcarbamoyl group and then reducing theN-methoxy-N-methylcarbamoyl group with lithium aluminium hydride.

Conversion from compound 29 into compound 30 can be performed byacetylating the secondary amino group of compound 29. Specifically,compound 29 can be reacted with an acetylating agent such as acetylchloride or acetic anhydride in a reaction solvent such as methylenechloride, THF or ethyl acetate in the presence of a base such as TEA,DIEA, pyridine or DMAP, for example.

Conversion from compound 30 into compound 31 can be performed bysimultaneously or stepwise eliminating the amino-protecting group P₇ andthe carboxyl-protecting group P₆ of compound 30. A specific deprotectingmethod comprises hydrogenolysis using a palladium catalyst when P₇ is abenzyloxycarbonyl group and P₆ is a benzyl group or treatment with anacid such as TFA when P₇ is a t-butoxycarbonyl group and P₆ is t-butylor alkali hydrolysis when P₆ is a methyl group.

Conversion from compound 31 into compound 32 can be performed byintramolecular cyclization of the amino group and the carboxyl group ofcompound 31 using a condensing reagent known to be useful for macrocycleformation reaction. Specifically, compound 32 can be obtained by, forexample, stirring compound 31 at room temperature under diluteconditions (at a concentration of compound 31 of about 0.005–0.02 M,preferably 0.01 M) using DMF, BOP and pyridine as reaction solvent,condensing agent and base, respectively.

When R₂′ of compound 32 is an optionally substituted protected aminogroup, conversion from compound 32 into compound 33 can be performed bydeprotecting the amino group, specifically hydrogenolysis using apalladium catalyst when the amino-protecting group of R₂′ is abenzyloxycarbonyl group or treatment with an acid such as TFA when it isa t-butoxycarbonyl group. When R₂′ is a hydrogen atom, compound 32 is adesired compound.

Alternatively, compounds of the present invention can be prepared byappropriately adapting the specific processes described in the examplesbelow.

EXAMPLES

The following examples further illustrate the present invention without,however, limiting the invention thereto. The chemical structuralformulae of the compounds of the examples are shown in Table A-1 andTable A-2 below.

TABLE A-1 Example No. Structural formula 1

2

3

4

5

6

7

8

9

10

In the following examples, ¹H-NMR and mass spectra were measured withthe following instruments. ¹H-NMR: JEOL JNM-EX-270 (270 MHz)

Mass spectra (FAB-MS): JASCO 70-250SEQ

Example 1 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide(1) Synthesis of Boc-Tyr(3-tert-Bu)-OH

To a mixed solution of 14.7 g (58.5 mmol) of H-Tyr(3-tert-Bu)-OMe and9.30 g (87.8 mmol) of sodium carbonate in 100 ml of 1,4-dioxane and 100ml of water was added 13.4 g (61.4 mmol) of di(tert-butyl) dicarbonatewith ice cooling and the mixture was stirred at room temperature for 1hour. The reaction solution was combined with water and extracted withethyl acetate, and then the extract was successively washed withsaturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over anhydrous magnesium sulfate,and then the solvent was distilled off under reduced pressure to give24.2 g of crude Boc-Tyr (3-tert-Bu)-OMe.

To a mixed solution of 16.5 g of the crude Boc-Tyr (3-tert-Bu)-OMe in120 ml of methanol and 40 ml of water was added 3.30 g (78.8 mmol) oflithium hydroxide monohydrate and the mixture was stirred at roomtemperature for 80 minutes. The reaction solution was neutralized with2N hydrochloric acid with ice cooling and then acidified with 10%aqueous citric acid. The reaction solution was extracted with methylenechloride and dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure to give 15.0 g (quant.) of the titlecompound.

(2) Synthesis of Boc-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 2.50 g (7.42 mmol) of Boc-Tyr (3-tert-Bu)-OH and 2.87 g(8.16 mmol) of H-β-Ala-OBzl p-toluenesulfonate in 22 ml of DMF wereadded 1.00 g (7.42 mmol) of HOBt, 0.848 ml (7.42 mmol) of NMM and 1.56 g(8.16 mmol) of WSCI with ice cooling, and the mixture was stirred atroom temperature for 150 minutes. The reaction solution was combinedwith water and extracted with ethyl acetate, and then the extract wassuccessively washed with saturated aqueous NH₄Cl solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure to give 3.57 g (97%) of Boc-Tyr(3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.38 (9H, s), 1.41 (9H, s), 2.40–2.58 (2H, m),2.84–3.00 (2H, m), 3.38–3.50 (2H, m), 4.14–4.26 (1H, m), 5.00 (1H, brs),5.09 (2H, s), 6.20–6.30 (1H, m), 6.54 (1H, d, J=8.2 Hz), 6.85 (1H, dd,J=1.7, 8.2 Hz), 7.03 (1H, d, J=1.7 Hz), 7.30–7.40 (5H, m).

(3) Synthesis of H-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 3.55 g (7.13 mmol) of Boc-Tyr (3-tert-Bu)-β-Ala-OBzl in30 ml of methylene chloride was added 20 ml of TFA and the mixture wasstirred at room temperature for 40 minutes. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with chloroform andthe extract was washed with saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressured to give 2.60 g (92%) of H-Tyr(3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.39 (9H, s), 2.58 (2H, t, J=6.3 Hz), 2.92 (1H, d,J=9.5 Hz), 3.13 (1H, dd, J=4.0, 13.9 Hz), 3.48–3.60 (3H, m), 5.14 (2H,s), 6.60 (1H, d, J=8.2 Hz), 6.88 (1H, dd, J=2.0, 8.2 Hz), 7.07 (1H, d,J=2.0 Hz), 7.30–7.40 (5H, m), 7.58–7.66 (1H, m).

(4) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 2.10 g (5.28 mmol) of H-Tyr (3-tert-Bu)-β-Ala-OBzl and3.19 g (5.54 mmol) of Boc-N-Me-Lys (Z)-OH dicyclohexylammonium salt in22 ml of DMF were added 713 mg (5.28 mmol) of HOBt, 0.60 ml (5.25 mmol)of NMM and 1.06 g (5.54 mmol) of WSCI with ice cooling, and the mixturewas stirred at room temperature for 2 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and then theextract was successively washed with 10% aqueous citric acid solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure to give 3.42 g(84%) of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.14–1.60 (5H, m), 1.37 (9H, s), 1.45 (9H, s),1.72–1.90 (1H, m), 2.40–2.54 (2H, m), 2.55 (3H, s), 2.86–3.02 (2H, m),3.06–3.22 (2H, m), 3.36–3.50 (2H, m), 4.34–4.56 (2H, m), 4.76–4.88 (1H,m), 5.09 (4H, s), 5.31 (1H, s), 6.30–6.60 (2H, m), 6.58 (1H, d, J=8.2Hz), 6.83 (1H, dd, J=1.9, 8.2 Hz), 7.04 (1H, d, J=1.9 Hz), 7.30–7.42(10H, m).

(5) Synthesis of N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 3.10 g (4.01 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl in 20 ml of methylene chloride was added 10 ml ofTFA and the mixture was stirred at room temperature for 90 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 2.59 g (96%) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 0.80–1.40 (6H, m), 1.38 (9H, s), 2.30 (3H, s), 2.56(2H, t, J=6.3 Hz), 2.70–3.24 (5H, m), 3.53 (2H, q, J=6.3 Hz), 4.48–4.60(1H, m), 4.80–4.90 (1H, m), 5.12 (2H, s), 5.15 (2H, s), 6.62–6.86 (3H,m), 7.04 (1H, d, J=1.3 Hz), 7.30–7.42 (10H, m).

(6) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 2.50 g (3.71 mmol) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl and 1.26 g (4.48 mmol) of Boc-Phe (4-F)-OH in18.5 ml of THF were added 1.14 g (4.45 mmol) of CMPI and 1.29 ml (9.28mmol) of TEA with ice cooling and the mixture was stirred at roomtemperature for 3 hours. The reaction solution was combined with waterand extracted with ethyl acetate, and then the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:hexane=1:1) to give3.48 g (100%) of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 0.80–1.40 (6H, m), 1.33, 1.36 and 1.39 (total 18H, s),2.17 and 2.61 (total 3H, s), 2.44–3.60 (10H, m), 4.30–4.94 (3H, m),5.01, 5.04 and 5.10 (total 4H, s), 5.20–5.48 (1H, m), 6.28–7.20 (10H,m), 7.30–7.42 (10H, m).

(7) Synthesis of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 2.72 g (2.90 mmol) of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl in 50 ml of methanol was added 270 mg of 10%palladium on carbon, and the mixture was stirred overnight at roomtemperature under a hydrogen atmosphere. After filtration, the filtratewas concentrated under reduced pressure to give 2.02 g (97%) of Boc-Phe(4-F)-N-Me-Lys-Tyr (3-tert-Bu)-β-Ala-OH.

To a mixed solution of 2.02 g (2.83 mmol) of the Boc-Phe(4-F)-N-Me-Lys-Tyr (3-tert-Bu)-β-Ala-OH in 140 ml of DMF and 140 ml ofpyridine was added 3.13 g (7.08 mmol) of BOP reagent and the mixture wasstirred at room temperature for 18 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and the extractwas successively washed with saturated aqueous NH₄Cl solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 1.37 g (69%) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

¹H-NMR (CDCl₃): δ 1.00–2.02 (6H, m), 1.33, 1.37 and 1.39 (total 18H, s),2.30 (3H, s), 2.24–3.86 (10H, m), 4.40–4.90 (4H, m), 5.38–5.60 (2H, m),6.00–6.20 (1H, m), 6.60–7.18 (7H, m), 6.54–7.22 (7H, m).

(8) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 1.25 g (1.79 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 20 ml of methylene chloride was added 10 ml of TFA and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol=20:1) to give 657 mg (61%) of the titlecompound.

FAB-MS (M⁺+1): 598. ¹H-NMR (DMSO-d): δ 0.60–1.92 (6H, m), 1.27 and 1.28(total 9H, s), 2.06–2.26 (2H, m), 2.54–2.90 (4H, m), 2.65 (3H, s),3.06–3.60 (4H, m), 3.62–4.94 (3H, m), 6.54–7.22 (7H, m), 7.50–8.50 (3H,m), 9.01 and 9.10 (total 1H, s).

Example 2 Synthesis of(2S-(2S.13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide(1) Synthesis of Boc-Phe (4-benzyloxy-3-tert-Bu)-OMe

A suspension of 7.5 g of the crude Boc-Tyr (3-tert-Bu)-OMe obtained inExample 1 (1) and 8.85 g.(64.0 mmol) of potassium carbonate in 45 ml ofDMF was stirred at room temperature for 2.5 hours, and then 4.6 ml (38.3mmol) of benzyl bromide was added. The mixture was stirred overnight andthen combined with water and extracted with ethyl acetate. The organiclayer was washed with saturated aqueous sodium chloride solution anddried over anhydrous magnesium sulfate and then concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:n-hexane=1:5) to give7.5 g (94%) of the title compound.

(2) Synthesis of Boc-N-Me-Phe (4-benzyloxy-3-tert-Bu)-OH

To a mixed solution of 7.5 g (17.0 mmol) of Boc-Phe(4-benzyloxy-3-tert-Bu)-OMe in 60 ml of methanol and 20 ml of water wasadded 1.07 g (25.5 mmol) of lithium hydroxide monohydrate, and themixture was stirred at room temperature for 1.5 hours. The reactionsolution was neutralized with 2N hydrochloric acid with ice cooling, andthen acidified with 10% aqueous citric acid. The reaction solution wasextracted with methylene chloride and dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure to give 6.8 g(93%) of crude Boc-Phe (4-benzyloxy-3-tert-Bu)-OH.

To a mixed solution of 6.2 g (14.5 mmol) of the crude compound in 30 mlof THF and 3 ml of DMF were added 7.2 ml (116 mmol) of methyl iodidewith ice cooling, then 1.74 g (43.5 mmol) of 60% sodium hydride. Themixture was stirred at room temperature for 21 hours, and then stirredwith 0.58 g (14.5 mmol) of 60% sodium hydride and 2.7 ml (43.5 mmol) ofmethyl iodide for 7 hours. The reaction solution was combined with waterwith ice cooling and washed with a mixed solution of n-hexane (150ml)-ether (50 ml). The aqueous layer was neutralized with 2Nhydrochloric acid with ice cooling, acidified with 10% aqueous citricacid solution, and then extracted with ethyl acetate. The extract wassuccessively washed with water and saturated aqueous sodium chloridesolution, and then dried over anhydrous magnesium sulfate andconcentrated under reduced pressure to give 5.0 g (78%) of a crudeproduct of the title compound.

(3) Synthesis of Boc-N-Me-Phe (4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

To a solution of 2.68 g (6.08 mmol) of the crude Boc-N-Me-Phe(4-benzyloxy-3-tert-Bu)-OH and 2.35 g (6.69 mmol) of H-β-Ala-OBzlp-toluenesulfonate in 20 ml of DMF were added 0.82 g (6.08 mmol) ofHOBt, 0.69 ml (6.08 mmol) of NMM and 1.28 g (6.69 mmol) of WSCI with icecooling, and the mixture was stirred overnight at room temperature. Thereaction solution was combined with water and then 10% aqueous citricacid solution, and then extracted with ethyl acetate. The organic layerwas washed with saturated aqueous NaHCO₃ solution and saturated aqueoussodium chloride solution and dried over magnesium sulfate, and then thesolvent was distilled off under reduced pressure to give 3.70 g (quant.)of the title compound.

(4) Synthesis of N-Me-Phe (4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

To a solution of 3.64 g (6.03 mmol) of Boc-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl in 20 ml of methylene chloride wasadded 10 ml of TFA, and the mixture was stirred at room temperature for45 minutes. The reaction solution was concentrated under reducedpressure, and the residue was neutralized with saturated aqueous NaHCO₃solution, and then extracted with methylene chloride and the extract waswashed with saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.02 g (99%) of the titlecompound.

(5) Synthesis of Boc-N-Me-Lys (Z)-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

To a solution of 3.00 g (5.96 mmol) of N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl, 4.12 g (7.16 mmol) of Boc-N-Me-Lys(Z)-OH dicyclohexylamine salt and 1.98 g (7.75 mmol) of CMPI in 30 ml ofTHF was added 2.91 ml (20.9 mmol) of TEA, and the mixture was stirredovernight at room temperature. The reaction solution was combined withwater and extracted with ethyl acetate, and the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:n-hexane=1:1) to give2.95 g (56%) of the title compound.

(6) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

To a solution of 2.90 g (3.30 mmol) of Boc-N-Me-Lys (Z)-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl in 20 ml of methylene chloride wasadded 10 ml of TFA, and the mixture was stirred at room temperature for45 minutes. The reaction solution was concentrated under reducedpressure, and the residue was neutralized with saturated aqueous NaHCO₃solution, and then extracted with methylene chloride. The organic layerwas dried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure to give 2.53 g (98%) of crude N-Me-Lys(Z)-N-Me-Phe (4-benzyloxy-tert-Bu)-β-Ala-OBzl.

To a solution of 2.50 g (3.21 mmol) of the crude compound, 1.00 g (3.53mmol) of Boc-Phe (4-F)-OH and 0.98 g (3.85 mmol) of CMPI in 16 ml of THFwas added 0.98 ml (7.06 mmol) of TEA with ice cooling, and the mixturewas stirred overnight at room temperature. The reaction solution wascombined with water and extracted with ethyl acetate, and the extractwas successively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and thenconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:hexane=2:1) to give 3.00 g (89%) of the title compound.

(7) Synthesis of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 2.72 g (2.60 mmol) of Boc-Phe (4-F)-pN-Me-Lys(Z)-N-Me-Phe (4-benzyloxy-3-tert-Bu)-β-Ala-OBzl in 50 ml of methanol wasadded 0.60 g of palladium hydroxide on carbon, and the mixture wasstirred overnight at room temperature under a hydrogen atmosphere. Afterthe reaction solution was filtered, the filtrate was concentrated underreduced pressure to give 1.98 g of crude Boc-Phe (4-F)-N-Me-Lys-N-Me-Tyr(3-tert-Bu)-β-Ala-OH.

To a mixed solution of 1.98 g (2.60 mmol) of the crude compound in 130ml of DMF and 130 ml of pyridine was added 3.45 g (7.80 mmol) of BOPreagent, and the mixture was stirred at room temperature for 26 hours.The reaction solution was concentrated under reduced pressure, combinedwith water and extracted with ethyl acetate. The organic layer wassuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution,and then dried over magnesium sulfate and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(developing solvent; methylene chloride:methanol:aqueousammonia=20:1:0.1) to give 0.66 g (36%, 2 steps) of the title compound.

(8) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 0.60 g (0.843 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 10 ml of methylene chloride was added 3 ml of TFA, and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution and extracted with methylenechloride. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas dissolved in 8 ml of methanol with heating. Crystals precipitated atroom temperature were filtered off to give 265 mg (51%) of the titlecompound.

FAB-MS (M⁺+1): 612. ¹H-NMR (DMSO-d): δ 1.0–1.5 (6H, m), 1.28 (9H, s),2.0–2.3 (3H, m), 2.4–2.7 (2H, m), 2.48 and 2.64 (total 3H, s), 2.9–3.1(4H, m), 3.5–3.7 (2H, m), 5.22 (1H, d, J=10.9 Hz), 5.33 (1H, t, J=7.3Hz), 6.5–6.7 (2H, m), 6.9–7.1 (4H, m), 7.1–7.2 (2H, m), 7.7–7.8 (1H, m),8.97 (1H, s).

Example 3

(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide(1) Synthesis of Boc-Tyr (3-tert-Bu)-Gly-OBzl

To a solution of 2.30 g (6.81 mmol) of Boc-Tyr (3-tert-Bu)-OH and 1.51 g(7.50 mmol) of H-Gly-OBzl hydrochloride in 20 ml of DMF were added 0.92g (6.81 mmol) of HOBt, 0.78 ml (6.81 mmol) of NMM and 1.44 g (7.50 mmol)of WSCI with ice cooling, and the mixture was stirred at roomtemperature for 105 minutes. The reaction solution was combined withwater and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (developing solvent; ethylacetate:n-hexane=1:2) to give 1.57 g (48%) of the title compound.

(2) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-Gly-OBzl

To a solution of 1.52 g (3.14 mmol) of Boc-Tyr (3-tert-Bu)-Gly-OBzl in25 ml of methylene chloride was added 8 ml of TFA, and the mixture wasstirred at room temperature for 75 minutes. The reaction solution wasconcentrated under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, and then extracted withmethylene chloride, and the extract was washed with saturated aqueoussodium chloride solution. The organic layer was dried over magnesiumsulfate, and then the solvent was distilled off under reduced pressureto give 1.19 g (98%) of crude H-Tyr (3-tert-Bu)-Gly-OBzl.

To a solution of 1.15 g (2.99 mmol) of the crude compound and 1.89 g(3.29 mmol) of Boc-N-Me-Lys (Z)-OH dicyclohexylamine salt in 20 ml ofDMF were added 0.40 g (2.99 mmol) of HOBt, 0.34 ml (2.99 mmol) of NMMand 0.63 g (3.29 mmol) of WSCI with ice cooling, and the mixture wasstirred at room temperature for 2.5 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and then theextract was successively washed with 10% aqueous citric acid solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (developing solvent;ethyl acetate:n-hexane=2:1) to give 2.16 g (95%) of the title compound.

(3) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr (3-tert-Bu)-Gly-OBzl

To a solution of 2.12 g (2.79 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-Gly-OBzl in 24 ml of methylene chloride was added 8 ml ofTFA, and the mixture was stirred at room temperature for 1 hour. Thereaction solution was concentrated under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution, and thenextracted with methylene chloride. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 1.81 g of crude N-Me-Lys (Z)-Tyr (3-tert-Bu)-Gly-OBzl.

To a solution of 1.79 g (2.71 mmol) of the crude compound, 0.84 g (2.98mmol) of Boc-Phe (4-F)-OH and 0.83 g (3.25 mmol) of CMPI in 13 ml of THFwas added 0.83 ml (5.96 mmol) of TEA with ice cooling, and the mixturewas stirred overnight at room temperature. The reaction solution wascombined with water and extracted with ethyl acetate, and then theextract was successively washed with 10% aqueous citric acid solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; methylenechloride:methanol:aqueous ammonia=50:1:0.05) to give 2.34 g (90%, 2steps) of the title compound.

(4) Synthesis of(2S-(2S,12S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 1.99 g (2.15 mmol) of Boc-Phe (4-F)-pN-Me-Lys (Z)-Tyr(3-tert-Bu)-Gly-OBzl in 40 ml of methanol was added 0.40 g of palladiumhydroxide on carbon, and the mixture was stirred overnight at roomtemperature under a hydrogen atmosphere. The reaction solution wasfiltered, and then the filtrate was concentrated under reduced pressureto give 1.60 g of crude Boc-Phe (4-F)-N-Me-Lys-Tyr (3-tert-Bu)-Gly-OH.

To a mixed solution of 1.60 g (2.15 mmol) of the crude compound in 100ml of DMF and 100 ml of pyridine was added 2.85 g (6.45 mmol) of BOPreagent, and the mixture was stirred at room temperature for 23 hours.The reaction solution was concentrated under reduced pressure, combinedwith water, and extracted with ethyl acetate. The organic layer wassuccessively washed with 10% aqueous citric acid solution and saturatedaqueous sodium chloride solution, and then dried over magnesium sulfate,and concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (developing solvent; methylenechloride:methanol:aqueous ammonia=25:1:0.05) to give 0.88 g (60%, 2steps) of the title compound.

(5) Synthesis of(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 0.86 g (1.26 mmol) of(2S-(2S,12S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 15 ml of methylene chloride was added 5 ml of TFA, and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure, and the residue was neutralizedwith aqueous ammonia and extracted with methylene chloride. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was heated with 4ml of methanol. After standing at room temperature, solids were filteredoff to give 474 mg (65%) of the title compound.

FAB-MS (M⁺+1): 584. ¹H-NMR (DMSO-d): δ 0.6–2.0 (6H, m), 1.30 (9H, m),2.4–3.0 (6H, m), 2.58 and 2.78 (total 3H, s), 3.2–3.5 (2H, m), 3.6–4.4(3.5H, m), 4.9–5.0 (0.5H, m), 6.6–7.2 (8H, m), 8.6–8.8 (1H, m), 8.87(0.5H, d, J=7.3 Hz), 9.00 and 9.05 (total 1H, s), 9.17 (0.5H, d, J=7.3Hz).

Example 4 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide(1) Synthesis of Boc-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

To a solution of 2.41 g (7.15 mmol) of Boc-Tyr (3-tert-Bu)-OH and 1.66 g(8.58 mmol) of N-Me-β-Ala-OBzl (Chem. Pharm. Bull., 31, 10, 1983,3553–3561) in 22 ml of DMF were added 966 mg (7.15 mmol) of HOBt, 0.817ml (7.15 mmol) of NMM and 1.51 g (7.87 mmol) of WSCI with ice cooling,and the mixture was stirred at room temperature for 150 minutes. Thereaction solution was combined with water and extracted with ethylacetate, and then the extract was washed with saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.52 g (96%) of Boc-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.34, 1.36, 1.37, 1.40 and 1.42 (total 18H, s),2.00–3.60 (9H, m), 4.66–4.80 (1H, m), 5.09 and 5.10 (total 2H, s),5.16–5.44 (2H, m), 6.40–7.04 (3H, m), 7.30–7.42 (5H, m).

(2) Synthesis of H-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

To a solution of 3.44 g (6.72 mmol) of Boc-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl in 30 ml of methylene chloride was added 10ml of TFA, and the mixture was stirred at room temperature for 50minutes. The reaction solution was distilled off under reduced pressure,and the residue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 2.70 g (97%) of H-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.37 (9H, s), 2.16–3.14 (4H, m), 2.73 and 2.84 (total3H, s), 3.40–3.70 (2H, m), 3.85 and 3.97 (total 1H, t, J=6.9 Hz), 5.08and 5.10 (total 2H, s), 6.53 and 6.57 (total 1H, d, J=8.3 Hz), 6.80 (1H,dd, J=1.3, 8.3 Hz), 6.99 (1H, d, J=1.3 Hz), 7.30–7.44 (5H, m).

(3) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

To a solution of 2.50 g (6.07 mmol) of H-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzland 3.67 g (6.37 mmol) of Boc-N-Me-Lys (Z)-OH dicyclohexylamine salt in18 ml of DMF were added 820 mg (6.07 mmol) of HOBt, 0.728 ml (6.37 mmol)of NMM and 1.28 g (6.68 mmol) of WSCI with ice cooling, and the mixturewas stirred at room temperature for 2 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and then theextract was successively washed with saturated aqueous NaHCO₃ solutionand saturated aqueous sodium chloride solution. The organic layer wasdried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure to give 4.17 g (87%) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.20–1.60 (5H, m), 1.36 (9H, s), 1.46 (9H, s),1.78–1.92 (1H, m), 2.10–2.94 (4H, m), 2.61, 2.71 and 2.82 (total 6H, s),3.17 (2H, q, J=6.6 Hz), 4.28–4.64 (2H, m), 4.40–5.48 (3H, m), 5.09 and5.10 (total 4H, s), 6.50 and 6.57 (total 1H, d, J=7.9–8.2 Hz), 6.68 (1H,brs), 6.80–6.90 (1H, m), 7.01 (1H, s), 7.28–7.42 (10H, m).

(4) Synthesis of N-Me-Lys (Z)-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

To a solution of 4.02 g (5.10 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl in 25 ml of methylene chloride was added 10ml of TFA, and the mixture was stirred at room temperature for 40minutes. The reaction solution was distilled off under reduced pressure,and the residue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 3.51 g (96%) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 0.90–1.04 (2H, m), 1.20–1.60 (4H, m), 1.38 (9H, s),2.30 (3H, s), 2.54–3.16 (7H, m), 2.81 and 2.89 (total 3H, s), 3.50–3.80(2H, m), 4.89 (1H, brs), 5.04–5.20 (1H, m), 5.11 and 5.13 (total 4H, s),6.60–7.08 (4H, m), 7.28–7.40 (10H, m).

(5) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl

To a solution of 3.40 g (4.94 mmol) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl and 1.47 g (5.19 mmol) of Boc-Phe (4-F)-OHin 25 ml of THF were added 1.39 g (5.43 mmol) of CMPI and 1.51 ml (10.9mmol) of TEA with ice cooling, and the mixture was stirred at roomtemperature for 2 hours. The reaction solution was combined with water,extracted with ethyl acetate and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:hexane=1:1) to give4.22 g (90%) of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 0.80–1.90 (6H, m), 1.35 and 1.37 (total 18H, s),2.36–3.66 (16H, m), 4.60–5.40 (5H, m), 5.08 and 5.11 (total 4H, s),6.30–7.22 (7H, m), 7.30–7.42 (10H, m).

(6) Synthesis of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 4.00 g (4.19 mmol) of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl in 84 ml of methanol was added 400 mg of 10%palladium on carbon, and the mixture was stirred overnight at roomtemperature under a hydrogen atmosphere. After filtration, the filtratewas concentrated under reduced pressure to give 3.05 g (100%) of Boc-Phe(4-F)-N-Me-Lys-Tyr (3-tert-Bu)-N-Me-β-Ala-OH.

To a mixed solution of 2.25 g (3.09 mmol) of the thus obtained Boc-Phe(4-F)-N-Me-Lys-Tyr (3-tert-Bu)-N-Me-β-Ala-OH in 150 ml of DMF and 150 mlof pyridine was added 4.10 g (9.27 mmol) of BOP reagent, and the mixturewas stirred at room temperature for 16 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and then theextract was successively washed with 10% aqueous CUSO₄ solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 1.32 g (60%) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

¹H-NMR (CDCl₃): δ 1.20–2.02 (6H, m), 1.30, 1.33, 1.37, 1.38 and 1.47(total 18H, s), 2.40–3.30 (16H, m), 3.60–3.78 (1H, m), 4.30–5.50 (4H,m), 6.50–7.40 (7H, m).

(7) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 1.20 g (1.69 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 15 ml of methylene chloride was added 5 ml of TFA, and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol:aqueous ammonia=20:1:0.1) to give 1.08 g(92%) of the title compound.

FAB-MS (M⁺+1): 612. ¹H-NMR (DMSO-d): δ 0.50–3.40 (16H, m), 1.29 (9H, s),2.74, 2.78 and 2.87 (total 6H, s), 3.77 and 4.42 (total 1H, t, J=6.5Hz), 4.02–4.16 (1H, m), 4.88–5.04 (1H, m), 6.61 (1H, d, J=8.0 Hz), 6.78(1H, dd, J=2.0, 8.0 Hz), 6.90–7.24 (4H, m), 7.40–7.56 (1H, m), 8.31 and9.07 (total 1H, s), 8.40 and 8.79 (total 1H, d, J=8.9–9.2 Hz).

Example 5 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide(1) Synthesis of Boc-N-Me-Lys (Me, Z)-OH

To a solution of 3.50 g (9.21 mmol) of Boc-Lys (Z)-OH in 86 ml of THFwere added 1.37 g (34.3 mmol) of 60% sodium hydride and 4.27 ml (68.6mmol) of methyl iodide with ice cooling, and the mixture was stirred atroom temperature for 15 hours. The reaction solution was combined with10% aqueous citric acid solution with ice cooling, extracted with ethylacetate and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; ethyl acetate:n-hexane:=1:1) to give 3.60 g (96%) ofBoc-N-Me-Lys (Me, Z)-OH.

¹H-NMR (CDCl₃): δ 1.20–2.02 (6H, m), 1.46 (9H, s), 2.79 (3H, s), 2.91(3H, s), 3.22–3.34 (2H, m), 4.34–4.70 (1H, m), 5.12 (2H, s), 7.30–7.42(5H, m).

(2) Synthesis of Boc-N-Me-Lys (Me, Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 2.50 g (6.28 mmol) of H-Tyr (3-tert-Bu)-P-Ala-OBzl and2.82 g (6.91 mmol) of Boc-N-Me-Lys (Me, Z)-OH in 18.8 ml of DMF wereadded 849 mg (6.28 mmol) of HOBt, 0.718 ml (6.28 mmol) of NMM and 1.32 g(6.91 mmol) of WSCI with ice cooling, and the mixture was stirred atroom temperature for 90 minutes. The reaction solution was combined withwater, extracted with ethyl acetate and the extract was washed with 10%aqueous citric acid solution, saturated aqueous NaHCO₃ solution andsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure to give 4.81 g (97%) of Boc-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 1.10–1.90 (6H, m), 1.37 (9H, s), 1.45 (9H, s),2.40–2.60 (5H, m), 2.80–3.00 (2H, m), 2.88 and 2.96 (total 3H, s),3.16–3.50 (4H, m), 4.30–4.58 (2H, m), 5.09 and 5.11 (total 4H, s),6.30–6.68 (3H, m), 6.83 (1H, dd, J=1.7, 7.9 Hz), 7.03 (1H, d, J=1.7 Hz),7.30–7.42 (10H, m).

(3) Synthesis of N-Me-Lys (Me, Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

To a solution of 4.48 g (5.69 mmol) of Boc-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl in 30 ml of methylene chloride was added 10 ml ofTFA, and the mixture was stirred at room temperature for 40 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 3.80 g (97%) of N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 0.70–1.60 (6H, m), 1.38 (9H, s), 2.29 (3H, s),2.50–2.62 (2H, m), 2.68–2.90 (3H, m), 2.88 and 2.96 (total 3H, s),3.14–3.60 (4H, m), 4.40–4.70 (1H, m), 5.12 and 5.18 (total 4H, s),6.60–6.90 (3H, m), 7.02 (1H, s), 7.30–7.42 (10H, m).

(4) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl

To a solution of 3.51 g (5.10 mmol) of N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl and 1.52 g (5.36 mmol) of Boc-Phe (4-F)-OH in25.5 ml of THF were added 1.43 g (5.61 mmol) of CMPI and 1.56 ml (11.2mmol) of TEA with ice cooling, and the mixture was stirred at roomtemperature for 90 minutes. The reaction solution was combined withwater and extracted with ethyl acetate, and then the extract was washedwith saturated aqueous sodium chloride solution. The organic layer wasdried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (developing solvent; ethyl acetate:hexane=1:1) togive 4.59 g (94%) of Boc-Phe (4-F)-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

¹H-NMR (CDCl₃): δ 0.90–1.40 (5H, m), 1.30, 1.36 and 1.39 (total 18H, s),1.76–1.92 (1H, m), 2.10–3.60 (16H, m), 4.40–4.80 (2H, m), 4.99, 5.11 and5.13 (total 4H, s), 5.00–5.60 (2H, m), 6.50–7.40 (18H, m).

(5) Synthesis of(2S-(2S,13S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 4.32 g (4.53 mmol) of Boc-Phe (4-F)-pN-Me-Lys (Me,Z)-Tyr (3-tert-Bu)-β-Ala-OBzl in 50 ml of methanol was added 400 mg of10% palladium on carbon, and the mixture was stirred at room temperaturefor 4 hours under a hydrogen atmosphere. After filtration, the filtratewas concentrated under reduced pressure to give 3.32 g (100%) of Boc-Phe(4-F)-N-Me-Lys (Me)-Tyr (3-tert-Bu)-β-Ala-OH.

To a mixed solution of 2.30 g (3.16 mmol) of the thus obtained Boc-Phe(4-F)-N-Me-Lys (Me)-Tyr (3-tert-Bu)-β-Ala-OH in 150 ml of DMF and 150 mlof pyridine was added 4.19 g (9.48 mmol) of BOP reagent, and the mixturewas stirred at room temperature for 16 hours. The reaction solution wascombined with water, extracted with ethyl acetate and the extract waswashed with saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 811 mg (36%) of(2S-(2S,13S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

¹H-NMR (CDCl₃): δ 0.80–2.00 (6H, m), 1.34, 1.37 and 1.40 (total 18H, s),2.30–3.40 (16H, m), 4.40–4.70 (2H, m), 5.16–5.44 (2H, m), 6.40–7.20 (9H,m).

(6) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 748 mg (1.05 mmol) of(2S-(2S,13S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 10 ml of methylene chloride was added 5 ml of TFA, and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol=20:1) to give 608 mg (95%) of the titlecompound.

FAB-MS (M⁺+1): 612. ¹H-NMR (DMSO-d): δ 0.60–1.90 (6H, m), 1.26 and 1.28(total 9H, s), 2.20–2.90 (12H, m), 3.20–4.40 (6H, m), 4.80–4.90 (1H, m),6.61 (1H, d, J=7.9 Hz), 6.70–7.60 (7H, m), 8.08–8.60 (1H, m), 9.01 and9.11 (total 1H, s).

Example 6 Synthesis of(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide(1) Synthesis of N-(Boc-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester

To a solution of 2.01 g (5.96 mmol) of Boc-Tyr (3-tert-Bu)-OH and 1.51 g(6.56 mmol) of 4-aminobutyric acid benzyl ester hydrochloride (Helv.Chim. Acta., 80, 1997, 1253) in 24 ml of DMF were added 805 mg (5.96mmol) of HOBt, 0.750 ml (6.56 mmol) of NMM and 1.26 g (6.57 mmol) ofWSCI with ice cooling, and the mixture was stirred at room temperaturefor 90 minutes. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 2.49 g (82%) ofN-(Boc-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester.

¹H-NMR (CDCl₃): δ 1.37 (9H, s), 1.41 (9H, s), 1.62–1.78 (2H, m), 2.26(2H, t, J=7.3 Hz), 2.82–3.30 (4H, m), 4.20 (1H, q, J=7.3 Hz), 4.97 (1H,brs), 5.10 (2H, s), 5.83 (1H, brs), 6.54 (1H, d, J=7.9 Hz), 6.86 (1H,dd, J=1.7, 7.9 Hz), 7.04 (1H, d, J=1.7 Hz), 7.30–7.42 (5H, m).

(2) Synthesis of N-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester

To a solution of 2.42 g (4.73 mmol) ofN-(Boc-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester in 20 ml ofmethylene chloride was added 5 ml of TFA, and the mixture was stirred atroom temperature for 1 hour. The reaction solution was distilled offunder reduced pressure, and the residue was neutralized with saturatedaqueous NaHCO₃ solution, extracted with chloroform and the extract waswashed with saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 1.94 g (100%) ofN-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester.

¹H-NMR (CDCl₃): δ 1.39 (9H, s), 1.85 (2H, tt, J=6.6, 7.6 Hz), 2.38 (2H,t, J=7.6 Hz), 2.60 (1H, dd, J=9.2, 13.8 Hz), 3.14 (1H, dd, J=3.9, 13.8Hz), 3.30 (2H, q, J=6.6 Hz), 3.53 (1H, dd, J=3.9, 9.2 Hz), 5.12 (2H, s),6.61 (1H, d, J=7.9 Hz), 6.88 (1H, dd, J=2.0, 7.9 Hz), 7.07 (1H, d, J=2.0Hz), 7.30–7.42 (5H, m).

(3) Synthesis ofN-(Boc-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester

To a solution of 1.89 g (4.59 mmol) ofN-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester and 2.78 g (4.82mmol) of Boc-N-Me-Lys (Z)-OH in 14 ml of DMF were added 620 mg (4.59mmol) of HOBt, 0.787 ml (6.89 mmol) of NMM and 968 mg (5.05 mmol) ofWSCI with ice cooling, and the mixture was stirred at room temperaturefor 90 minutes. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.41 g (90%) ofN-(Boc-N-Me-Ne-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester.

¹H-NMR (CDCl₃): δ 1.12–1.90 (8H, m), 1.36 (9H, s), 1.44 (9H, s), 2.27(2H, t, J=7.3 Hz), 2.56 (3H, s), 2.90–3.30 (6H, m), 4.37 (1H, t, J=7.1Hz), 4.69 (1H, q, J=7.6 Hz), 4.82 (1H, brs), 5.09 (4H, s), 5.50–5.70 and6.00–6.20 (total 1H, m), 6.47 (1H, d, J=4.3 Hz), 6.59 (1H, d, J=7.9 Hz),6.84 (1H, dd, J=1.7, 7.9 Hz), 7.04 (1H, d, J=1.7 Hz), 7.30–7.44 (10H,m).

(4) Synthesis of N-(N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester

To a solution of 3.32 g (4.21 mmol) ofN-(Boc-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester in 20 ml of methylene chloride was added 10 ml of TFA, and themixture was stirred at room temperature for 40 minutes. The reactionsolution was distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure to give2.86 g (99%) of N-(N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

¹H-NMR (CDCl₃): δ 0.80–0.98 (2H, m), 1.10–1.70 (4H, m), 1.38 (9H, s),1.81 (2H, tt, J=6.6, 7.3 Hz), 2.28 (3H, s), 2.35 (2H, t, J=7.3 Hz),2.76–3.20 (5H, m), 3.27 (2H, q, J=6.6 Hz), 4.46–4.58 (1H, m), 4.80–4.94(1H, m), 5.11 and 5.14 (total 4H, s), 6.40–6.52 (1H, m), 6.69 (1H, d,J=7.9 Hz), 6.84 (1H, dd, J=1.6, 7.9 Hz), 6.88 (1H, brs), 7.04 (1H, d,J=1.6 Hz), 7.30–7.46 (10H, m).

(5) Synthesis ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester

To a solution of 2.82 g (4.10 mmol) ofN-(N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl esterand 1.28 g (4.51 mmol) of Boc-Phe (4-F)-OH in 21 ml of THF were added1.26 g (4.92 mmol) of CMPI and 1.14 ml (8.20 mmol) of TEA with icecooling, and the mixture was stirred at room temperature for 2 hours.The reaction solution was combined with water and extracted with ethylacetate, and then the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; ethyl acetate:hexane=1:1) to give 3.57 g (91%) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

¹H-NMR (CDCl₃): δ 0.80–1.50 (6H, m), 1.35, 1.37 and 1.38 (total 18H, s),1.70–1.86 (2H, m), 2.15 and 2.62 (total 3H, s), 2.24–2.38 (2H, m),2.60–3.30 (8H, m), 4.28–4.94 (3H, m), 5.26–5.40 (1H, m), 6.08–7.20 (10H,m), 7.30–7.42 (10H, m).

(6) Synthesis of(2S-(2S,14S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 3.50 g (3.67 mmol) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester in 100 ml of methanol was added 400 mg of 10%palladium on carbon, and the mixture was stirred overnight at roomtemperature under a hydrogen atmosphere. After filtration, the filtratewas concentrated under reduced pressure to give 2.65 g (99%) ofN-(Boc-4-fluorophenylalanyl-N-Me-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid.

To a mixed solution of 2.35 g (3.22 mmol) of the thus obtainedN-(Boc-4-fluorophenylalanyl-N-Me-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid in 160 ml of DMF and 160 ml of pyridine was added 4.27 g (9.66mmol) of BOP reagent, and the mixture was stirred at room temperaturefor 16 hours. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 1.88 g (82%) of(2S-(2S,14S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

¹H-NMR (CDCl₃): δ 1.04–1.50 (6H, m), 1.29, 1.34 and 1.39 (total 18H, s),1.70–1.94 (2H, m), 2.10–2.40 (2H, m), 2.24 and 2.55 (total 3H, s),2.56–3.70 (8H, m), 4.40–4.94 (3H, m), 5.24–5.40 (1H, m), 5.80–6.44 (2H,m), 6.62–7.20 (7H, m), 8.21 (1H, d, J=8.2 Hz).

(7) Synthesis of(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 1.75 g (2.46 mmol) of(2S-(2S,14S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 20 ml of methylene chloride was added 10 ml of TFA, and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol:aqueous ammonia=20:1:0.1) to give 783 mg(52%) of the title compound.

FAB-MS (M⁺+1): 612. ¹H-NMR (DMSO-d): δ 0.60–2.00 (6H, m), 1.06 (2H, t,J=6.9 Hz), 1.28 and 1.29 (total 9H, s), 2.06–2.22 (2H, m), 2.50–3.00(9H, m), 3.20–4.04 (3H, m), 4.24–4.44 (3/2H, m), 4.96 (1/2H, d, J=9.9Hz), 6.61 (1H, d, J=8.3 Hz), 6.70–6.84 (1H, m); 6.90–7.74 (7H, m), 8.14and 8.95 (total 1H, d, J=7.6–8.2 Hz), 9.04 and 9.09 (total 1H, s).

Example 7 Synthesis of(2S,14S)-13-(2S-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane(1) Synthesis of Z-Tyr (3-tert-Bu)-β-Ala-OtBu

To a solution of 2.69 g (7.25 mmol) of Z-Tyr (3-tert-Bu)-OH and 1.45 g(7.98 mmol) of H-β-Ala-OtBu hydrochloride in 22 ml of DMF were added 980mg (7.25 mmol) of HOBt, 1.66 ml (14.5 mmol) of NMM and 1.53 g (7.98mmol) of WSCI with ice cooling, and the mixture was stirred at roomtemperature for 90 minutes. The reaction solution was combined withwater and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure to give 2.76 g (76%) of Z-Tyr(3-tert-Bu)-β-Ala-OtBu.

¹H-NMR (CDCl₃): δ 1.37 (9H, s), 1.41 (9H, s), 2.33 (2H, q, J=6.3 Hz),2.88–3.06 (2H, m), 3.40 (2H, q, J=6.3 Hz), 4.20–4.34 (1H, m), 4.80–5.00(1H, m), 5.09. (2H, s), 5.20–5.30 (1H, m), 6.12–6.24 (1H, m), 6.57 (1H,d, J=7.9 Hz), 6.86 (1H, d, J=7.9 Hz), 7.03 (1H, s), 7.30–7.42 (5H, m).

(2) Synthesis of H-Tyr (3-tert-Bu)-β-Ala-OtBu

To a solution of 2.70 g (5.42 mmol) of Z-Tyr (3-tert-Bu)-β-Ala-OtBu in80 ml of methanol was added 300 mg of 10% palladium on carbon, and themixture was stirred at room temperature for 5 hours under a hydrogenatmosphere. After filtration, the filtrate was concentrated underreduced pressure to give 1.96 g (99%) of H-Tyr (3-tert-Bu)-p-Ala-OtBu.

¹H-NMR (CDCl₃): δ 1.36 (9H, s), 1.43 (9H, s), 2.40 (2H, t, J=6.6 Hz),2.77 (1H, dd, J=8.2, 13.5 Hz), 3.14 (1H, dd, J=4.6, 13.5 Hz), 3.36–3.50(2H, m), 3.76–3.84 (1H, m), 6.71 (1H, d, J=7.9 Hz), 6.93 (1H, d, J=7.9Hz), 7.04 (1H, s), 7.64–7.72 (1H, m).

(3) Synthesis of Z-N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-OtBu

To a solution of 1.90 g (5.22 mmol) of H-Tyr (3-tert-Bu)-β-Ala-OtBu and2.18 g (5.74 mmol) of Z-N-Me-Lys (Boc)-OH in 17.5 ml of DMF were added705 mg (5.22 mmol) of HOBt, 0.656 ml (5.74 mmol) of NMM and 1.10 g (8.14mmol) of WSCI with ice cooling, and the mixture was stirred at roomtemperature for 1 hour. The reaction solution was combined with waterand extracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.49 g (92%) of Z-N-Me-Lys(Boc)-Tyr (3-tert-Bu)-β-Ala-OtBu.

¹H-NMR (CDCl₃): δ 0.90–1.10 (2H, m), 1.20–1.50 (4H, m), 1.37 (9H, s),1.43 (9H, s), 1.44 (9H, s), 2.34–2.46 (2H, m), 2.80 (1H, dd, J=9.2, 14.2Hz), 2.99 (2H, q, J=6.6 Hz), 3.10–3.24 (1H, m), 3.43 (2H, q, J=6.0 Hz),3.94–4.04 (1H, m), 4.55 (1H, q, J=6.2 Hz), 4.65 (1H, brs), 5.10 (2H, s),5.20–5.30 (1H, m), 6.34 (1H, d, J=6.0 Hz), 6.42 (1H, brs), 6.60 (1 h,brs), 6.66 (1H, d, J=7.9 Hz), 6.82 (1H, dd, J=1.7, 7.9 Hz), 7.01 (1H, d,J=1.7 Hz), 7.30–7.42 (5H, m).

(4) Synthesis of(2S,14S)-13-amino-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

To a solution of 2.51 g (3.46 mmol) of Z-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-OtBu in 10 ml of methylene chloride was added 10 ml ofTFA, and the mixture was stirred at room temperature for 50 minutes. Thereaction solution was distilled off under reduced pressure, and a mixedsolution of the residue in 170 ml of acetonitrile and 170 ml of pyridinewas stirred with 4.59 g (10.4 mmol) of BOP reagent at room temperaturefor 21 hours. The solvent was distilled off under reduced pressure, andthe residue was successively washed with diethyl ether and chloroform togive solids. The thus obtained solids were dissolved in 50 ml ofmethanol and combined with 300 mg of 10% palladium on carbon, and themixture was stirred overnight at room temperature under a hydrogenatmosphere. After filtration, the filtrate was concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; chloroform:methanol aqueousammonia=10:1:0.1) to give 1.30 g (90%) of(2S,14S)-13-amino-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane.

¹H-NMR (CD₃OD): δ 0.90–1.40 (6H, m), 1.40 (9H, s), 2.28–2.50 (2H, m),2.78 (1H, dd, J=7.6, 13.9 Hz), 3.06 (2H, dd, J=6.9, 13.5 Hz), 3.30–3.72(4H, m), 4.65 (1H, t, J=7.6 Hz), 6.66 (1H, d, J=7.9 Hz), 6.90 (1H, dd,J=1.7, 7.9 Hz), 7.08 (1H, d, J=1.7 Hz).

(5) Synthesis of(2S,14S)-13-((2S)-2-(tert-butoxycarbonylamino)-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

To a solution of 0.89 g (2.13 mmol) of(2S,14S)-13-amino-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecaneand 739 mg (2.77 mmol) of Boc-Phe (4-F)-H in 22 ml of methanol wereadded 1 ml of acetic acid and 423 mg (6.39 mmol) of 95% sodiumcyanoborohydride with ice cooling, and the mixture was stirred at roomtemperature for 30 minutes. The reaction solution was combined withwater and extracted with chloroform, and then the extract was washedwith saturated aqueous sodium chloride solution. The organic layer wasdried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (developing solvent; chloroform:methanol:aqueousammonia=20:1:0.1) to give 793 mg (56%) of(2S,14S)-13-((2S)-2-(tert-butoxycarbonylamino)-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane.

¹H-NMR (CDCl₃): δ 1.12–1.80 (6H, m), 2.06 (1H, dd, J=5.3, 11.9 Hz), 2.18(1H, dd, J=6.3, 11.9 Hz), 2.28–2.64 (3H, m), 2.70–2.80 (2H, m),2.94–3.12 (3H, m), 3.42–3.68 (3H, m), 4.72 (1H, dd, J=6.3, 9.3 Hz), 6.65(1H, d, J=7.9 Hz), 6.88 (1H, dd, J=2.0, 7.9 Hz), 7.00 (2 h, t, J=8.2Hz), 7.11 (1H, d, J=2.0 Hz), 7.21 (2H, dd, J=7.6, 8.2 Hz).

(6) Synthesis of(2S,14S)-13-((2S)-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

To a solution of 735 mg (1.10 mmol) of(2S,14S)-13-((2S)-2-(tert-butoxycarbonylamino)-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecanein 5 ml of methylene chloride was added 3 ml of TFA, and the mixture wasstirred at room temperature for 40 minutes. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with chloroform andthe extract was washed with saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (developing solvent;chloroform:methanol:aqueous ammonia=5:1:0.1) to give 608 mg (97%) of thetitle compound.

FAB-MS (M⁺+1): 570. ¹H-NMR (DMSO-d): δ 1.00–1.52 (4H, m), 1.30 (9H, s),1.64–1.94 (4H, m), 2.14–2.32 (3H, m), 2.50–2.90 (5H, m), 3.20–3.62 (4H,m), 4.42–4.56 (1H, m), 6.63 (1H, d, J=8.2 Hz), 6.77 (1H, d, J=8.2 Hz),6.93 (1H, s), 7.00–7.24 (5H, m), 7.69 (1H, d, J=5.9 Hz), 8.09 (1H, d,J=9.2 Hz), 9.00–9.20 (1H, m).

Example 8 Synthesis of(2S-(2S.13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-N-methyl-1,4,8-triaza-3,14-dioxocyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)propionamide(1) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-OMe

To a solution of 2.00 g (7.97 mmol) of H-Tyr (3-tert-Bu)-OMe and 5.05 g(8.77 mmol) of Boc-N-Me-Lys (Z)-OH in 24 ml of DMF were added 1.08 g(7.97 mmol) of HOBt, 1.37 ml (12.0 mmol) of NMM and 1.68 g (8.77 mmol)of WSCI with ice cooling, and the mixture was stirred at roomtemperature for 3 hours. The reaction solution was combined with waterand extracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:hexane 1:1) to give 4.99 g (100%) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-OMe.

¹H-NMR (CDCl₃): δ 1.14–1.30 (2H, m), 1.37 (9H, s), 1.43 (9H, s),1.40–1.64 (3H, m), 1.78–1.96 (1H, m), 2.56 (3H, s), 2.93 (1H, dd, J=7.3,14.2 Hz), 3.04–3.22 (3H, m), 3.73 (3H, s), 4.44 (1H, t, J=7.6 Hz),4.72–4.86 (2H, m), 5.09 (2H, s), 5.55 (1H, brs), 6.40 (1H, brs), 6.61(1H, d, J=7.9 Hz), 6.77 (1 h, d, J=7.9 Hz), 6.95 (1H, s), 7.30–7.40 (5H,m).

(2) Synthesis of Boc-N-Me-Lys-Tyr (3-tert-Bu)-OMe

To a solution of 4.92 g (7.85 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-OMe in 80 ml of methanol was added 500 mg of 10% palladiumon carbon, and the mixture was stirred overnight at room temperatureunder a hydrogen atmosphere. After filtration, the filtrate wasconcentrated under reduced pressure to give 3.87 g (100%) ofBoc-N-Me-Lys-Tyr (3-tert-Bu)-OMe.

¹H-NMR (CDCl₃): δ 1.20–1.94 (6H, m), 1.36 (9H, s), 1.43 (9H, s), 2.63(3H, s), 2.80–3.00 (3H, m), 3.12 (1H, dd, J=5.0, 13.8 Hz), 3.74 (3H, s),4.45 (1H, t, J=6.9 Hz), 4.70–4.80 (1H, m), 6.73 (1H, d, J=7.9 Hz), 6.82(1H, d, J=7.9 Hz), 6.94 (1H, s).

(3) Synthesis of Boc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe

To a solution of 3.10 g (6.29 mmol) of Boc-N-Me-Lys-Tyr (3-tert-Bu)-OMeand 1.43 g (6.92 mmol) of N-phenylmethoxycarbonyl-3-aminopropanal in 63ml of methanol were added 0.36 ml of acetic acid and 832 mg (12.6 mmol)of 95% sodium cyanoborohydride with ice cooling, and the mixture wasstirred at room temperature overnight. The reaction solution wascombined with water and extracted with chloroform, and then the extractwas washed with saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol:aqueous ammonia=20:1:0.1) to give 2.30 g (53%) ofBoc-N-Me-Ne-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr (3-tert-Bu)-OMe.

¹H-NMR (CDCl₃): δ 1.16–2.00 (8H, m), 1.37 (9H, s), 1.44 (9H, s), 2.61(3H, s), 2.72 (2H, t, J=7.3 Hz), 2.85 (2H, t, J=6.6 Hz), 2.84–2.94 (1H,m), 3.12 (1H, dd, J=5.3, 13.9 Hz), 3.24–3.38 (2H, m), 3.74 (3H, s), 4.42(1H, t, J=7.3 Hz), 4.75 (1H, q, J=6.6 Hz), 5.10 (2H, s), 5.40–5.50 (1H,m), 6.56–6.66 (1H, m), 6.70 (1H, d, J=7.9 Hz), 6.78 (1H, dd, J=1.7, 7.9Hz), 6.95 (1H, d, J=1.7 Hz), 7.28–7.40 (5H, m).

(4) Synthesis ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Ne-acetyl-Lys-Tyr(3-tert-Bu)-OMe

To a solution of 2.24 g (3.27 mmol) ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr (3-tert-Bu)-OMe in33 ml of methylene chloride were added 0.401 ml (4.25 mmol) of aceticanhydride and 0.684 ml (4.91 mmol) of TEA with ice cooling, and themixture was stirred for 30 minutes. The reaction solution wasneutralized with saturated aqueous NaHCO₃ solution, extracted withmethylene chloride and the extract was washed with saturated aqueoussodium chloride solution. The organic layer was dried over magnesiumsulfate, and then the solvent was distilled off under reduced pressure,and the residue was purified by silica gel column chromatography(developing solvent; chloroform:methanol:aqueous ammonia=30:1:0.1) togive 1.92 g (81%) ofBoc-N-Me-Ne-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe.

¹H-NMR (CDCl₃): δ 1.08–2.00 (8H, m), 1.37 (9H, s), 1.43 (9H, s), 2.05(3.H, s), 2.96 (1H, dd, J=6.9, 14.5 Hz), 3.04–3.42 (7H, m), 3.73 (3H,s), 4.30–4.60 (1H, m), 4.76 (1H, q, J=6.3 Hz), 5.09 (2H, s), 5.77 (1H,brs), 6.30–6.50 (1H, m), 6.58–6.82 (2H, m), 6.95 (1H, s), 7.30–7.40 (5H,m).

(5) Synthesis ofN-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe

To a solution of 1.82 g (2.50 mmol) ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Nε-acetyl-Lys-Tyr(3-tert-Bu)-OMe in 20 ml of methylene chloride was added 10 ml of TFA,and the mixture was stirred at room temperature for 40 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 1.56 g (99%) ofN-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe.

¹H-NMR (CDCl₃): δ 0.74–1.82 (8H, m), 1.38 and 1.39 (total 9H, s), 2.02and 2.07 (total 3H, s), 2.35 and 2.37 (total 3H, s), 2.60–3.40 (9H, m),3.76 and 3.77 (total 3H, s), 4.78–5.00 (3/2H, m), 5.12 (2H, s),5.52–5.60 (1/2H, m), 6.60–7.04 (7/2H, m), 7.30–7.46 (5H, m), 7.67 (1/2H,m).

(6) Synthesis of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe

To a solution of 1.50 g (2.39 mmol) ofN-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe and 811 mg (2.87 mmol) of Boc-Phe (4-F)-OH in 16 ml ofTHF were added 794 mg (3.11 mmol) of CMPI and 0.733 ml (5.26 mmol) ofTEA with ice cooling, and the mixture was stirred at room temperaturefor 14 hours. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:hexane=2:1) to give1.96 g (92%) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe.

¹H-NMR (CDCl₃): δ 1.00–1.90 (8H, m), 1.35, 1.36 and 1.38 (total 18H, s),2.04 (3H, s), 2.20 and 2.55 (total 3H, s), 2.66–3.42 (10H, m), 3.70 and3.76 (total 3H, s), 4.60–5.04 (3H, m), 5.10 (2H, s), 5.20–6.24 (3H, m),6.58–7.20 (7H, m), 7.30–7.44 (5H, m).

(7) Synthesis of Boc-Phe (4-F)-N-Me-Nε-acetyl-Nε-(3-aminopropyl)-Lys-Tyr(3-tert-Bu)-OH

To a solution of 1.89 g (2.12 mmol) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe in 20 ml of dioxane was added 2 ml of 2N aqueous sodiumhydroxide solution, and the mixture was stirred at room temperature for100 minutes. The reaction solution was combined with 10% aqueous citricacid solution with ice cooling, extracted with chloroform and theextract was washed with saturated aqueous sodium chloride solution. Theorganic layer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 1.87 g (100%) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OH.

To a solution of 1.82 g (2.08 mmol) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OH in 20 ml of methanol was added 200 mg of 10% palladium oncarbon, and the mixture was stirred overnight at room temperature undera hydrogen atmosphere. After filtration, the filtrate was concentratedunder reduced pressure to give 1.32 g (85%) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-aminopropyl)-Lys-Tyr (3-tert-Bu)-OH.

¹H-NMR (DMSO-d): δ 1.00–2.00 (8H, m), 1.28 and 1.38 (total 18H, s), 1.96(3H, s), 2.30–3.50 (9H, m), 4.00–5.00 (3H, m), 6.54–6.98 (3H, m),7.00–7.30 (4H, m), 7.30–7.44 (1H, m), 8.96 (1H, d, J=5.3 Hz).

(8) Synthesis of(2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-tert-butoxycarbonylamino-3-(4-fluorophenyl)-N-methylpropionamide

To a mixed solution of 1.10 g (1.48 mmol) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-aminopropyl)-Lys-Tyr (3-tert-Bu)-OH in 75 mlof DMF and 75 ml of pyridine was added 1.96 g (4.44 mmol) of BOPreagent, and the mixture was stirred at room temperature for 18 hours.The reaction solution was concentrated under reduced pressure, combinedwith water and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution and saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; methylene chloride:methanol:aqueousammonia=20:1:0.05) to give 0.59 g (55%) of2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-tert-butoxycarbonylamino-3-(4-fluorophenyl)-N-methylpropionamide.

(9) Synthesis of(2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 0.55 g (0.757 mmol) of(2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-tert-butoxycarbonylamino-3-(4-fluorophenyl)-N-methylpropionamidein 7 ml of methylene chloride was added 3 ml of TFA, and the mixture wasstirred at room temperature for 55 minutes. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with methylenechloride and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; methylene chloride:methanol:aqueous ammonia=15:1:0.1) to give0.43 g (90%) of the title compound.

FAB-MS (M⁺+1): 626. ¹H-NMR (DMSO-d6-CDCl₃): δ 0.7–1.8 (16H, m), 1.29 and1.30 (total 9H, s), 1.9–2.0 (3H, m), 2.4–3.4 (10H, m), 2.62 and 2.64(3H, s), 3.70 (0.5H, m), 4.0–4.4 (2H, m), 4.92 (0.5H, d, J=9.2 Hz),6.5–7.2 (8H, m), 7.8–8.1 (1.5H, m), 9.0–9.1 (1.5H, m).

Example 9 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9,N-dimethyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)propionamide(1) Synthesis of Boc-N-Me-Orn (Me) (Z)-OH

To a solution of 3.00 g (8.19 mmol) of Boc-Orn (Z)-OH in 82 ml of THFwere added 1.31 g (32.8 mmol) of 60% sodium hydride and 4.08 ml (65.5mmol) of methyl iodide with ice cooling, and the mixture was stirred atroom temperature for 16 hours. The reaction solution was combined withwater with ice cooling, and washed with hexane. The aqueous layer wascombined with 10% aqueous citric acid solution, extracted with ethylacetate and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure to give3.22 g (100%) of Boc-N-Me-Orn (Me) (Z)-OH.

¹H-NMR (CDCl₃): δ 1.46 (9H, s), 1.50–2.02 (4H, m), 2.70–2.90 (3H, m),2.92 (3H, s), 3.20–3.46 (2H, m), 4.32–4.80 (1H, m), 5.13 (2H, s),7.24–7.40 (5H, m).

(2) Synthesis ofN-(Boc-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester

To a solution of 2.39 g (5.80 mmol) ofN-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester and 2.51 g (6.38mmol) of Boc-N-Me-Orn (Z)-OH in 29 ml of DMF were added 784 mg (5.80mmol) of HOBt, 0.729 ml (6.38 mmol) of NMM and 1.22 g (6.38 mmol) ofWSCI with ice cooling, and the mixture was stirred at room temperaturefor 140 minutes. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:hexane=1:1) to give 3.91 g (86%) ofN-(Boc-N-Me-Ne-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester.

¹H-NMR (CDCl₃): δ 1.36 (9H, s), 1.45 (9H, s), 1.50–1.90 (4H, m), 2.26(2H, t, J=7.3 Hz), 2.47 and 2.58 (total 3H, s), 2.88 (3H, s), 2.95 (2H,d, J=7.3 Hz), 3.10–3.44 (4H, m), 4.42–4.60 (2H, m), 5.09 (2H, s), 5.12(2H, s), 5.30–5.50 (1H, m), 5.90–6.10 (1H, m), 6.54 (1H, d, J=7.9 Hz),6.83 (1H, d, J=7.9 Hz), 7.04 (1H, s), 7.24–7.40 (10H, m).

(3) Synthesis of N-(N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester

To a solution of 3.81 g (4.84 mmol) ofN-(Boc-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester in 40 ml of methylene chloride was added 10 ml of TFA, and themixture was stirred at room temperature for 40 minutes. The reactionsolution was distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure to give3.26 g (98%) of N-(N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

¹H-NMR (CDCl₃): δ 1.06–1.40 (2H, m), 1.37 (9H, s), 1.78 (2H, tt, J=6.9,7.3 Hz), 2.27 (3H, s), 2.32 (2H, t, J=7.3 Hz), 2.76–2.92 (5H, m),2.98–3.30 (3H, m), 3.225 (2H, q, J=6.9 Hz), 4.34–4.56 (1H, m), 5.10 and5.14 (total 4H, s), 6.30–6.88 (3 h, m), 7.02 (1H, d, J=2.6 Hz),7.30–7.62 (11H, m).

(4) Synthesis ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester

To a solution of 3.10 g (4.51 mmol) ofN-(N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester and 1.41 g (4.96 mmol) of Boc-Phe (4-F)-OH in 45 ml of THF wereadded 1.38 g (5.41 mmol) of CMPI and 1.38 ml (9.92 mmol) of TEA with icecooling, and the mixture was stirred at room temperature for 14 hours.The reaction solution was combined with water and extracted with ethylacetate, and then the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; ethyl acetate:hexane=2:1) to give 4.20 g (98%) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

¹H-NMR (CDCl₃): δ 1.20–1.50 (2H, m), 1.34 and 1.38 (total 18H, s),1.60–1.84 (4H, m), 2.04–3.40 (16H, m), 4.30–5.06 (5/2H, m), 5.08 and5.11 (total 4H, s), 5.29 (1/2H, d, J=6.6 Hz), 5.60–6.30 (2H, m),6.40–7.20 (7H, m), 7.24–7.42 (21/2H, m), 7.68–7.76 (1/2H, m).

(5)(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 2.80 g (2.68 mmol) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester in 40 ml of methanol was added 0.52 g of palladiumhydroxide on carbon, and the mixture was stirred at room temperature for5 hours under a hydrogen atmosphere. The reaction solution was filtered,and the filtrate was concentrated under reduced pressure, and a mixedsolution of 2.27 g of the residue in 135 ml of DMF and 135 ml ofpyridine was stirred with 3.56 g (8.04 mmol) of BOP reagent at roomtemperature for 12 hours. The reaction solution was concentrated underreduced pressure, combined with water and extracted with ethyl acetate,and then the extract was successively washed with 10% aqueous citricacid solution and saturated aqueous sodium chloride solution. Theorganic layer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; methylenechloride:methanol:aqueous ammonia=20:1:0.1) to give 1.14 g (60%) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

(6) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

To a solution of 1.10 g (1.54 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 9 ml of methylene chloride was added 3 ml of TFA, and the mixture wasstirred at room temperature for 1 hour. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with methylenechloride and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; methylene chloride:methanol:aqueous ammonia=13:1:0.1) to give609 mg (65%) of the title compound.

FAB-MS (M⁺+1): 612. ¹H-NMR (DMSO-d6-CDCl₃): δ 0.8–2.0 (6H, m), 1.29 (9H,s), 2.0–3.4 (12H, m), 3.6–4.5 (3.5H, m), 4.9–5.1 (0.75H, m), 5.4–5.7(0.75H, m), 6.6–7.3 (10H, m), 7.6–7.8 (1H, m), 8.0–8.3 (1H, m), 8.9–9.2(2H, m).

Example 10 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide(1) Synthesis of H-Tyr (3-tert-Bu)-β-Ala-O-tert-Bu

To a mixed solution of 3.5 g (13.9 mmol) of H-Tyr (3-tert-Bu)-OMe and2.21 g (20.9 mmol) of sodium carbonate in 25 ml of 1,4-dioxane and 25 mlof water was added 2.09 ml (14.6 mmol) of benzyloxycarbonyl chloridewith ice cooling, and the mixture was stirred at room temperature for 2hours. The reaction solution was combined with water, extracted withethyl ether, and then the extract was washed with saturated aqueoussodium chloride solution. The organic layer was dried over anhydrousmagnesium sulfate, and then the solvent was distilled off under reducedpressure, and the resulting crude Z-Tyr (3-tert-Bu)-OMe in 20 ml of1,4-dioxane was stirred with a solution of 1.05 g (26.3 mmol) of sodiumhydroxide in 10 ml of water at room temperature for 40 minutes. Thereaction solution was washed with ethyl ether, and then the aqueouslayer was acidified with concentrated hydrochloric acid and extractedwith methylene chloride. The extract was dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure, and 1.88 g (13.9mmol) of HOBt, 1.6 ml (13.9 mmol) of NMM and 2.93 g (15.3 mmol) of WSCIwere added to a solution of the resulting crude Z-Tyr (3-tert-Bu)-OH and2.78 g (15.3 mmol) of H-β-Ala-O-tert-Bu hydrochloride in 40 ml of DMFwith ice cooling, and the mixture was stirred overnight at roomtemperature. The reaction solution was combined with water, 10% aqueouscitric acid solution and extracted with ethyl acetate, and then theextract was successively washed with saturated aqueous NaHCO₃ solutionand saturated aqueous sodium chloride solution. The extract was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure, and a solution of the resulting crude Z-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu in 70 ml of methanol was stirred with 1.0 gof 20% palladium hydroxide on carbon for 1 hour under a hydrogenatmosphere. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure to give 4.70 g (93%) of the titlecompound.

¹H-NMR (CDCl₃): δ 1.39 (9H, s), 1.45 (9H, s), 2.44 (2H, t, J=6.3 Hz),2.57 (1H, dd, J=9.4, 13.9 Hz), 3.17 (1H, dd, J=4.0, 13.9 Hz), 3.47–3.56(3H, m), 6.64 (1H, d, J=7.9 Hz), 6.90 (1H, dd, J=2.0, 7.9 Hz), 7.08 (1H,d, J=2.0 Hz), 7.67 (1H, brt).

(2) Synthesis of Fmoc-N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-O-tert-Bu

To a solution of 3.97 g (10.9 mmol) of H-Tyr (3-tert-Bu)-β-Ala-O-tert-Buand 5.26 g (10.9 mmol) of Fmoc-N-Me-Lys (Boc)-OH in 40 ml of DMF wereadded 1.47 g (10.9 mmol) of HOBt, 1.25 ml (10.9 mmol) of NMM and 2.09 g(10.9 mmol) of WSCI with ice cooling, and the mixture was stirredovernight at room temperature. The reaction solution was combined withwater and 10% aqueous citric acid solution and extracted with ethylacetate, and then the extract was successively washed with saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (using ethyl acetate:n-hexane=1:1 aseluent) to give 8.14 g (90%) of the title compound.

¹H-NMR (CDCl₃): δ 1.0–1.9 (6H, m), 1.34 (9H, s), 1.42 (9H, s), 1.44 (9H,s), 2.3–2.4 (2H, m), 2.55 (3H, brs), 2.7–2.9 (1H, m), 3.0–3.1 (3H, m),3.4–3.5 (2H, m), 4.2–4.6 (6H, m), 6.45 (2H, brs), 6.75 (1H, brs), 7.00(1H, s), 7.3–7.5 (5H, m), 7.58 (2H, d, J=6.3 Hz), 7.79 (2H, d, J=7.6Hz).

(3) Synthesis of N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-O-tert-Bu

To a solution of 6.8 g (8.20 mmol) of Fmoc-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu in 15 ml of methylene chloride was added 15ml of diethylamine, and the mixture was stirred at room temperature for95 minutes. The reaction solution was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (using methylene chloride:methanol aqueousammonia=30:1:0.1 as eluent) to give 4.57 g (91%) of the title compound.

¹H-NMR (CDCl₃): δ 0.8–0.9 (2H, m), 1.0–1.7 (4H, m), 1.38 (9H, s), 1.44(9H, s), 1.48 (9H, s), 2.33 (3H, s), 2.43 (2H, t, J=5.9–6.3 Hz), 2.7–3.1(4H, m), 3.25 (1H, dd, J=4.6, 14.2 Hz), 3.49 (2H, dd, J=5.9–6.3,11.9–12.2 Hz), 4.5–4.7 (2H, m), 6.7–6.8 (2H, m), 6.86 (1H, d, J=6.6 Hz),7.03 (1H, s), 7.38 (1H, d, J=7.9 Hz).

(4) Synthesis of Z-Phe-N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-O-tert-Bu

To a solution of 4.57 g (7.53 mmol) of N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu, 2.93 g (9.79 mmol) of Z-Phe-OH and 2.69 g(10.5 mmol) of CMPI in 20 ml of THF was added 3.1 ml (22.6 mmol) of TEAwith ice cooling, and the mixture was stirred overnight at roomtemperature. The reaction solution was combined with water and extractedwith ethyl acetate, and then the extract was successively washed withsaturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (developing solvent;ethyl acetate:hexane=3:1) to give 6.7 g (quant.) of the title compound.

(5) Synthesis of(2S-(2S,13S))-2-benzyloxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide

To a solution of 6.7 g (7.53 mmol) of Z-Phe-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu in 40 ml of methylene chloride was added 40ml of TFA, and the mixture was stirred at room temperature for 70minutes. The reaction solution was concentrated under reduced pressure,and the residue was combined with toluene, and the solvent was distilledoff under reduced pressure. A mixed solution of the resulting crudeZ-Phe-N-Me-Lys-Tyr (3-tert-Bu)-β-Ala-OH in 380 ml of DMF and 380 ml ofpyridine was stirred with 9.99 g (22.6 mmol) of BOP reagent overnight atroom temperature. The reaction solution was concentrated under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; chloroform methanol:aqueousammonia=80-20:1:0.05). The residue was dissolved in ethyl acetate, andsuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution,and dried over anhydrous magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 2.6 g (48%) of the titlecompound.

(6) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide

To a solution of 1.25 g (1.79 mmol) of(2S-(2S,13S))-2-benzyloxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamidein 35 ml of methanol was added 0.55 g of 10% palladium hydroxide oncarbon, and the mixture was stirred for 2.5 hours under a hydrogenatmosphere. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol:aqueous ammonia=20:1:0.1) to give 1.48 g (73%) ofthe title compound.

FAB-MS (M⁺+1): 580

¹H-NMR (DMSO-d6-CDCl₃): δ 0.60–2.0 (6H, m), 1.29 (9H, s), 2.1–2.3 (2H,m), 2.5–2.9 (4H, m), 2.66 and 2.60 (3H, s), 3.1–3.3 (2H, m), 3.5–3.6(2H, m), 3.71 and 3.92 (1H, t, J=6.3 Hz), 4.18 and 4.94 (1H, d, J=11.9Hz), 4.4–4.6 (1H, m), 6.60–6.64 (1H, m), 6.72 and 6.75 (1H, dd, J=1.9,8.0 Hz), 6.86 and 6.93 (1H, d, J=1.9 Hz), 7.05–7.29 (5H, m), 7.6–7.8(2H, m), 8.09 and 8.39 (1H, d, J=8.8–9.4), 8.9–9.1 (1H, s).

In the following test examples, representative compounds of the presentinvention were evaluated for motilin receptor antagonist activity bypharmacological tests in order to demonstrate the utility of compoundsof the present invention.

Test Example 1 Motilin Receptor Binding Test

Motilin receptor binding activities of compounds of the presentinvention were tested as follows [Vantrappen et al., Regul. Peptides,15, 143 (1986)].

A protein solution was prepared by homogenizing the duodenum isolatedfrom a killed rabbit in 50 mM Tris after stripping the mucosa. Theprotein solution was incubated with 25 pM ¹²⁵I motilin for 2 hours at25° C., and then the radioactivity bound to the protein was measuredwith a γ-counter (COBRA II™, model 5005, Packard). The differencebetween the radioactivity of the protein solution incubated alone andthe radioactivity of the protein solution incubated with a large excessof motilin (10⁻⁷ M) was recorded as specific binding of motilin tomotilin receptors.

The concentration required to inhibit 50% of the specific binding ofmotilin to motilin receptors (IC₅₀, nM) was determined for the compoundssynthesized in Examples 1–10 above. As a comparative example, the IC₅₀of the compound of formula (5) below, i.e. a cyclic peptide derivativehaving motilin antagonist activity (the compound described inJP-A-7–138284) was also determined. The results are shown in Table B-1.

Test Example 2 Effect on the Contraction of a Sample of the LongitudinalDuodenal Muscle Extracted from Rabbit

The effect of compounds of the present invention on motilin-inducedcontraction of a sample of the longitudinal muscle of the duodenumextracted from a rabbit was tested as follows.

A duodenal specimen (5×15 mm) extracted from a killed rabbit wassuspended in the direction of the longitudinal muscle in an incubator at28° C. filled with Krebs solution (10 ml organ bath). A mixed gas (95%O₂, 5% CO₂) was continuously passed through the Krebs solution and thecontraction of the duodenal specimen was isotonically recorded (under aload of 1 g) via an isotonic transducer (ME-3407, ME Commercial, Tokyo,Japan). The extent of contraction of the duodenal specimen was expressedas a percentage to the contraction induced by acetylcholine at aconcentration of 10⁻⁴ M.

The effect of the compounds synthesized in Examples 1–8 and 10 on theconcentration-dependent contraction induced by motilin added into theincubator was determined to calculate pA₂ (see “Drug receptors” editedby Takayanagi, published by Nanzando). The results are shown in TableB-1. As a comparative example, the results of the compound of formula(5) are also shown.

TABLE B-1 Motilin receptor Contraction binding test inhibition testExample No. IC₅₀ (nM) pA₂ 1 1.2 9.0 2 1.4 8.9 3 0.52 8.2 4 0.78 9.0 51.4 9.9 6 0.87 8.5 7 4.5 8.0 8 0.84 8.2 9 5.3 10  2.2 7.9 Comparative 107.2 example

Table B-1 shows that the compounds of the present invention haveexcellent motilin receptor antagonist activity as compared with thecompound of the comparative example.

INDUSTRIAL APPLICABILITY

Compounds of the present invention have motilin receptor antagonistactivity or the like and are useful as pharmaceuticals such aspharmaceuticals for treating irritable bowel syndrome.

1. A compound of general formula (1):

wherein R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle; R₂ represents a hydrogen atom or anoptionally substituted amino group; R₃ represents a hydrogen atom, amethyl group or an ethyl group; R₄ represents a hydrogen atom, a methylgroup or an ethyl group; R₅ represents a hydrogen atom, a methyl groupor an ethyl group; R₆ represents a hydrogen atom, a methyl group, anethyl group or —CO—R₇; R₇ represents a hydrogen atom or a straight orbranched alkyl group having 1–3 carbon atoms; V represents a carbonylgroup or a methylene group; W represents a carbonyl group or a methylenegroup: X represents a carbonyl group or a methylene group; Y representsa carbonyl group or a methylene group; Z represents a carbonyl group ora methylene group; m represents a number of 0–2; and n represents anumber of 0–3; except for the case where R₁ represents a phenyl group,R₂ represents an amino group, all of R₃, R₄, R₅ and R₆ represent ahydrogen atom, V represents a methylene group, all of W, X, Y and Zrepresent a carbonyl group, and both m and n represent 1; or a hydrateor a pharmaceutically acceptable salt thereof.
 2. The compound ofgeneral formula (1) of claim 1 wherein X is a methylene group when R₁ isa phenyl group and R₃ is a hydrogen atom, or a hydrate or apharmaceutically acceptable salt thereof.
 3. The compound of generalformula (1) of claim 1, wherein R₁ is a phenyl group or ahalogen-substituted phenyl group, or a hydrate or a pharmaceuticallyacceptable salt thereof.
 4. The compound of general formula (1) of anyone of claim 1, wherein R₂ is an amino group, or a hydrate or apharmaceutically acceptable salt thereof.
 5. The compound of generalformula (1) of claim 1, wherein R₃ is a hydrogen atom or a methyl group,or a hydrate or a pharmaceutically acceptable salt thereof.
 6. Thecompound of general formula (1) of claim 1, wherein R₄ is a hydrogenatom or a methyl group, or a hydrate or a pharmaceutically acceptablesalt thereof.
 7. The compound of general formula (1) of claim 1, whereinR₅ is a hydrogen atom or a methyl group, or a hydrate or apharmaceutically acceptable salt thereof.
 8. The compound of generalformula (1) of claim 1, wherein R₆ is a hydrogen atom, a methyl group oran acetyl group, or a hydrate or a pharmaceutically acceptable saltthereof.
 9. The compound of general formula (1) of claim 1, wherein V isa methylene group, or a hydrate or a pharmaceutically acceptable saltthereof.
 10. The compound of general formula (1) of claim 1, wherein Yis a carbonyl group, or a hydrate or a pharmaceutically acceptable saltthereof.
 11. The compound of general formula (1) of claim 1, wherein Zis a carbonyl group, or a hydrate or a pharmaceutically acceptable saltthereof.
 12. The compound of general formula (1) of claim 1 wherein R₁is a phenyl group or a halogen-substituted phenyl group, R₂ is an aminogroup, R₃ is a hydrogen atom or a methyl group, R₄ is a hydrogen atom ora methyl group, R₅ is a hydrogen atom or a methyl group, R₆ is ahydrogen atom, a methyl group or an acetyl group, V is a methylenegroup, W is a carbonyl group or a methylene group, X is a carbonyl groupor a methylene group, Y is a carbonyl group and Z is a carbonyl group,or a hydrate or a pharmaceutically acceptable salt thereof.
 13. Thecompound of claim 1 selected from the group consisting of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S,14S)-13-(2S-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane,(2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-N-methyl-1,4,8-triaza-3,14-dioxocyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)propionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9,N-dimethyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)propionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide,or a hydrate or a pharmaceutically acceptable salt thereof.
 14. Apharmaceutical comprising the compound of claim 1, or a hydrate or apharmaceutically acceptable salt thereof as an active ingredient.
 15. Amotilin receptor antagonist comprising the compound of claim 1, or ahydrate or a pharmaceutically acceptable salt thereof.
 16. Agastrointestinal motility inhibitor comprising the compound of claim 1,or a hydrate or a pharmaceutically acceptable salt thereof as an activeingredient.
 17. A pharmaceutical for treating hypermotilinemiacomprising the compound of claim 1, or a hydrate or a pharmaceuticallyacceptable salt thereof as an active ingredient.
 18. A compound ofgeneral formula (2):

wherein R₃ represents a hydrogen atom, a methyl group or an ethyl group;R₄ represents a hydrogen atom, a methyl group or an ethyl group; R₅represents a hydrogen atom, a methyl group or an ethyl group; R₆represents a hydrogen atom, a methyl group, an ethyl group or —CO—R₇; mrepresents a number of 0–2; n represents a number of 0–3; P₂ representsa hydrogen atom or a protective group for a phenolic hydroxyl group; P₃represents a hydrogen atom or a protective group for a carboxyl group;P₄ represents a hydrogen atom or a protective group for an amino group;and P₅ represents a hydrogen atom or a protective group for an aminogroup; or a hydrate or a pharmaceutically acceptable salt thereof.
 19. Acompound of general formula (3):

wherein R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle; R₃ represents a hydrogen atom, amethyl group or an ethyl group; R₄ represents a hydrogen atom, a methylgroup or an ethyl group; R₅ represents a hydrogen atom, a methyl groupor an ethyl group; m represents a number of 0–2; n represents a numberof 0–3; P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group; P₃ represents a hydrogen atom or a protectivegroup for a carboxyl group; P₅ represents a hydrogen atom or aprotective group for an amino group; and R₂′ represents a hydrogen atomor an optionally substituted protected amino group; or a hydrate or apharmaceutically acceptable salt thereof.
 20. A compound of generalformula (4):

wherein R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle; R₃ represents a hydrogen atom, amethyl group or an ethyl group; R₄ represents a hydrogen atom, a methylgroup or an ethyl group; R₅ represents a hydrogen atom, a methyl groupor an ethyl group; R₂′ represents a hydrogen atom or an optionallysubstituted protected amino group; m represents a number of 0–2; nrepresents a number of 0–3; P₆ represents a hydrogen atom or aprotective group for a carboxyl group; and P₇ represents a hydrogen atomor a protective group for an amino group; or a hydrate or apharmaceutically acceptable salt thereof.
 21. A method for treatinghypermotilinemia comprising the step of administering a therapeuticallyeffective amount of the compound of claim 1, or a hydrate or apharmaceutically acceptable salt thereof to a patient in need of suchtreatment.